text:
A Masters degree gives you the opportunity to either further your understanding of a particular question or take off in a completely different way using skills you have gained from your previous undergraduate degree.
Business studies is an academic program that is designed for students wanting to learn more about how businesses are run. Courses may include more general topics or more specific ones to help prepare students for a particular career.
Taiwan, officially the Republic of China, is a state in East Asia and the first constitutional republic in Asia. Taiwan has several good universities, many of which have exchange agreements with foreign universities, and these are a good way to experience life in Taiwan.
Top Master Programs in Business Studies in Taiwan 2017
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Amid knowledge economy era, more and more successful enterprises have already shifted organization resource from mass production to unique design and innovation. For enhancing market competition, increasing sales volume and profitability and avoiding low-price war, Taiwan government and enterprises highly aware have already advocated the concept of design value added aggressively. Taiwan design industry has won 450 awards in 4 major international design awards in Japan G-Mark, USA IDEA, Germany iF and Reddot, including 10 top awards since 2003. Taiwan industry design even won 65 awards in Germany iF design competition in 2006, surpassing Japan and Korean and ranking the second next to host country Germany and topping in Asia. It proves Taiwan designers have already reached top international level and also "design" is a trend that Taiwan must continue development. ... [-]


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Five Spice Marinade
Total Time
Prep 20 mins
Cook 20 mins
A chinese-style marinade. Recommended to use with pork, but might be good with chicken? Prep time does not include marinating time.
Ingredients Nutrition
- 1 lb pork loin, partially frozen, then sliced into thick strips
- 1⁄4 cup soy sauce
- 1⁄4 cup hoisin sauce
- 1⁄4 cup dry white wine
- 2 garlic cloves, minced
- 1⁄4 teaspoon salt
- 1 teaspoon five-spice powder
- 2 tablespoons sugar
- Mix all marinade ingredients.
- Marinate pork strips six hours, turning occasionally.
- Drain; discard marinade.
- Grill pork.
- Note: I have never actually timed how long it takes to cook the pork, I just make sure it is done, so the cooking time may be off.
Most Helpful
This is a great marinade. We used it to make chicken wings. I marinated the wings for about 8 hours and then baked them at 350 degrees for 45-50 minutes. I did broil them for about 1 minute just to get them extra brown. I really like five-spice-powder! I doubled this recipe and it was great! Thanks ~Lainie~! We will make this again!

subset: commoncrawl


text:
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Marcus Redpath
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|Marshal Marcus Redpath
|Undead (Humanoid)
| Alliance (formerly)
The Forsaken (formerly, unvillingly), Rotbrain
|Rotbrain Encampment, Tirisfal Glades
|Killable; Undead
|Joseph, Carlin, Marlene (presumed)
Marcus Redpath (aka Marshal Redpath) was a human quest giver located in Southshore in the contested territory of the Hillsbrad Foothills. He seemed to be the leader of the military forces in Southshore and was eager to rid the town of various threats, including the murlocs, Crushridge ogres, and naga.
CataclysmEditThis section concerns content exclusive to Cataclysm.
With the destruction of Southshore by Forsaken plague bombing in Cataclysm, Marcus was killed with many other citizens. His body was transported to the Deathknell Graves and raised as an undead, but when he was approached by a Forsaken agent to join them, he refused, vowing vengeance against them for destroying Southshore.
He is later found at Rotbrain Encampment as the leader of the Rotbrain undead, planning an attack on Deathknell, where Forsaken players are sent to slay him.
Objective of Edit
- [1] The Wakening (to talk to)
- [5] Assault on the Rotbrain Encampment
Removed quests EditNote: These quests are no longer associated with this particular mob/NPC/item.
When human in Southshore, he started the following quests:
[32] Farren's Proof
[36] Crushridge Bounty
[40] Crushridge Warmongers (Elite)
Background EditThis section concerns content exclusive to the Warcraft novels or short stories.
Marcus Redpath was the headman of Southshore prior to the onset of the Second War, greeting Anduin Lothar, Khadgar, and the refugees from the Kingdom of Stormwind. He sent his fastest rider, Willem, to King Terenas to alert him to their arrival.
He is possibly related to Joseph, Carlin, and Marlene.
He might have been the Redpath briefly mentioned in the quest Daelin's Men, which would have made him a member of the Theramore city guard at some point.
Patch changes Edit
- Patch 4.0.3 (15-Nov-2010): Killed and raised as undead.
- Patch 1.1.0 (07-Nov-2004): Added.


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text: Well when I first installed bootcamp with windows XP I was able to access the root hard drive from OS X and copy across certain gaming files to places like my documents. However I needed to up the hard drive capacity so I copied across the program files and documents and settings files, removed the partition and made a larger one and reinstalled it. However now I am not able to copy the files back across to the windows partition whatever I try to do... please help! I want my games back!


text:
- WiFi:
- Microwave:
- Refrigerator:
- Hair Dryer:
- Iron/Ironing Board:
- Coffee/Tea Maker:
Hyatt Place Delray Beach is located in the Pineapple Grove Arts District of downtown Delray Beach within 1 mile of the Gold Coast and its beautiful beaches and walking distance to Atlantic Avenue shopping, dining and entertainment districts. Conveniently accessible from I-95, the hotel is located near PBIA, FLL and Fort Lauderdale Cruise Port.
Enjoy a $10 Food & Beverage credit per day. Valid at the Lobby Bar and Coffee to Cocktails bar. Includes complimentary parking. View details


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Black Friday, otherwise known as the Friday after Thanksgiving, is the time when retailers slash prices on anything and everything from electronics to baby accessories. Over the past few years, retailers have... read more
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Two issues were recently raised in the Carrollton City Council, and both are disturbing.
First a councilman and councilwoman both argued for weakening enforcement of laws which mandate that citizens clean up their yards, and don't park unlicensed junk cars on their property. While both council members are fine people, they are just wrong on this issue.
Last summer when the city hired Art Zook as a code enforcement officer, I took a tour of the city and found dozens of yards filled with trash and junk. Frankly, I was ashamed at the accumulated junk and blight imposed on the whole community by a few people who felt no shame in blighting their community.
Recently, I took another tour of Carrollton to see if any progress has been made since the city hired Mr. Zook and directed him to inspect code violations and seek enforcement of the laws. I was pleasantly surprised when I noticed a vast improvement. Literally dozens of yards had been cleaned up. Junk cars were gone, and a lot of yards had removed trash and debris.
We should remember that Zook isn't making up the rules, he was hired to seek enforcement of the existing zoning laws. Those who want a blighted city should seek to change the law and not attack the officer who is merely enforcing the law.
The second issue raised at the council meeting was the chauvinist argument that Zook was a newcomer to Carrollton. He has lived in Carrollton for six years and is married to a lifelong resident.
The council members mentioned are suggesting that he shouldn't be supported in his work of cleaning up our neighborhoods "because he wasn't born here."
These arguments are attempts to suggest that citizens who have moved here from somewhere else should not merit the respect and credibility of the City Council and are second-class citizens. This is an effort to divert our attention away from the yards full of trash and junked cars and to try to focus the issue on "outsiders."
Every current doctor, lawyer and dentist in Carrollton moved here from somewhere else. Several members of the City Council and many city employees have moved here. I represented this county in the state Legislature and in the District Court, and I moved here from somewhere else.
Former Mayor Charlie Webster presided over a period of the most productive economic development of the city, and he moved to Carrollton from Northern Kentucky. I know of at least two other former mayors who moved here from somewhere else.
We should be thankful for the new blood that has enlivened our community and who have chosen to bring their families here, to open businesses here, to employ people here, and to share the tax burden. People who move to Carrollton have made the lives of everyone here better. I would suspect that there are many registered voters in Carrollton who have moved here from somewhere else who will remember these slights at the next city election.
If there are two classes of citizenship in Carrollton, will "outsiders" like myself get a refund on our property taxes?
Let's not be distracted. The argument isn't about Enforcement Officer Zook's length of residence here, it's about a few council members who believe arguing for a blighted community will generate votes at the next election.
Judge Stan Billingsley, retired
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107 Pickering Ln Charleston SC 29414 MLS# 1028735
1028735 - Residential real estate
Charleston, SC
Beautiful 2 bedroom, 2 full bath condo in Grand Oaks. Very spacious floor plan with an over-sized family room for all the entertaining one could want. Family room also contains a gas fireplace for those cold winter nights. Screened porch located off family room which backs up to a nice wooded area, very quiet setting. Smooth cathedral ceilings as soon as you enter the front door along with beautiful crown molding throughout. The Ashford plan has two master suite size bedrooms which are approximately 15'x18' and 11.5'x14'. One master bedroom contains two closets for both his and hers. The master bathroom comes equipped with a garden tub and separate shower with both his and her vanities. This is not your average Condo! Community pool for just Grove Park residents. Possible short sale at listing price.
View the Full Details of this Home for Sale in Charleston
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Everyone has a wish list, I suppose—a list of things they hope to do before they die. I have one, too. It's several pages. . . read more
As an art tutor (and artist) I know that self-doubt and confidence is one of the biggest things for artists to overcome. These. . . read more
If you like art—as a professional, student, or teacher—your source of fresh ideas is somewhere out in the world. As Rumi said: "What you seek is seeking you." It's a matter of connecting.
So today I'd like to share three ways to find artistic inspiration, and give you an easy tool to capture those sources of inspiration to. . . read more
Art is a communication form, yet most of us don't know what we are trying to communicate and need to try lots of different. . . read more
Life is hard. . . art can be hard. And making a living with art? Well, that can feel impossible. I know; I've been there. A. . . read more
Have you ever felt discouraged about asking for things? Maybe you sometimes feel that you're going to be rejected anyway. . . so why bother? I know I've often felt that way, but time and experience has taught me to get back on the horse and try again.
In fact, one of the best things I've learned over the last few years as an independent artist (and writer) is how important it is. . . read more
Starting something is easy. Finishing? Not so much. It's difficult enough to finish things when they go relatively swiftly, but. . . read more
Artists, compared to the general population, are known to have unique traits. One of these is curiosity. If you find that. . . read more
You ever notice how the best products for artists are almost always made by other artists? How many times have you needed something in the studio and said, "Why can't someone just invent ____________? I'd use it!"
Well, Stickybones fits that scenario exactly.
. . . read more
"Human rights and the widespread denial of human rights is the most important subject in the world. At our peril, we close. . . read more
At one point or another, I think everyone feels like their time is being stretched. Work, family, random events you can't get. . . read more


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Kassius Holdorf was out for revenge against Maki Pitolo at Victory FC 54's headlining affair. And, after a thunderous knockout in the early seconds of the second round, he managed to not only avenge read news >>
UFC welterweight Kyle Noke announced his retirement via Twitter following his decision loss to Omari Akhmedov at UFC Fight Night 101. read news >>
Jeff Curran returned to the cage for the first time since March 2015 to face Raufeon Stots at Victory FC 53. Stots did not provide a warm welcome back to the cage.Filed under: News read news >>
UFC veteran Manny Gamburyan has called it quits on his mixed martial arts career. read news >>
MMA fighter Miesha Tate announced her retirement from the UFC on Saturday evening. "It's not my time anymore," she announced, per SportsCenter, after losing a unanimous decision to Raquel Pennington read news >>
Raquel Pennington won a unanimous decision and Miesha Tate called it a career. read news >>
Image via YouTube The UFC has become more and more reliant on the small number of superstars they have on their roster, the two most prominent being Conor McGregor and Ronda Rousey. Internal UFC docu read news >>
Vitor Belfort's historic mixed martial arts career has come to an end after his technical-knockout loss against Gegard Mousasi at UFC 204 on Oct. 8. Belfort announced his decision to hang up the glov read news >>
In advance of UFC 204, Dan Henderson was adamant he would retire regardless of how his rematch with Michael Bisping proceeded. As it turns out, per Ariel Helwani of MMAFighting.com, Henderson stayed read news >>
Legacy FC 60 headliner Tyler East announced his retirement from MMA after rival Brian Heden knocked him out. read news >>
It was fun while it lasted, but Floyd Mayweather put the Conor McGregor hype train out of service. read news >>
In a sport that never stops moving, even MMA junkies can struggle to keep up with all the latest news. If you missed any of Monday's biggest headlines, catch up with "The Comeback."Filed under: Bella read news >>
Brazilian jiu-jitsu ace Rafael Lovato toppled fellow submission ace Marcelo Nunes, scoring a second round TKO to claim Legacy FC's vacant middleweight title.Filed under: AXS TV Fights, News read news >>
The UFC is still trying to figure out what to do with Conor McGregor. The former UFC 200 main eventer lost his spot on the card Tuesday after ominously tweeting that he wanted "to retire young" and, read news >>


text:
Ricotta -Spinach Casserole
Total Time
Prep 15 mins
Cook 40 mins
One way to get them to eat spinach!
Ingredients Nutrition
- 6 eggs
- 1 1⁄2 cups ricotta cheese
- 2 tablespoons instant minced onion
- 2 tablespoons flour
- salt
- 1⁄2 teaspoon dry mustard
- 1 (10 ounce) package frozen chopped spinach, cook drained and well squeezed
- Beat all the ingredients (except spinach) together and mix until well blended Stir in the spinach.
- Heat oven to 350 degrees F.
- Grease a 1 1/2 quart casserole dish.
- Pour in the spinach mixture.
- Bake apprx 35 minutes or until a knife inserted in the middle comes out clean.
Most Helpful
This surpassed my expectations. I didn't think a spinach dish could be this good! We don't have 10 ounce packs of frozen spinach but I used a 250 gram (or 8.8 ounce) pack with great results. To reduce the fat I removed 3 of the egg yolks from the recipe, using 3 whites and 3 whole eggs. I don't feel this hurt the flavour of the recipe at all. Omitted the mustard as I didn't have any. Also, silly me forgot to grease the baking dish so it did stick a bit around the edges, so don't forget this step like I did! Great recipe which everyone loved, thanks Bergy.
Bergy this was wonderfull, did a half recipe for breakfast this morning. Mmmmmmmmmmm Mmmmmmmm!! great combination of flavours, ricotta cheese, onion and spinach! we will do this one often, thanks for sharing!
I made this for company last night and everyone loved this dish. Thank you for sharing.


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Gender: Boy
Meaning: One with pale white complexion
Origin: Hindi, Indian, Malayali
Religion: Hindu
Panduranga is a Hindu Boy name, meaning of the name is One with pale white complexion and origin of the name is Hindi, Indian, Malayali. This name is mostly used in Hindu.
If you select the name Panduranga for your baby, Congratulations! But if you don't like Panduranga then please continue searching our site.
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Business development, develop relationship with business heads, technology leaders at various level.
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7 to 8 years experience of sales in IT services. Good orientation of IBM technology would be preferred. Working with captive clients would be an added advantage.
Experience: 7 to 10 years
It Sales Business Development It Services Sales Manager Sales It Sales Manager Pricing


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Browse | Recently Added | Popular | Buy/RequestView My Bounty to keep track of what you have, want, and like
What is Bounty?Bounty is an exclusive database tracking all official Boba Fett collectibles. We have 1,784 items documented.
|Release Year
|Dark Horse Comics (re-print)
|The classic adaptation of the second Star Wars movie continues as Luke's Jedi training with Yoda progresses, and the Rebels' fight against the Empire takes a turn for the worse as Han Solo gets himself into a rather sticky situation. This second of two volumes reprints the Marvel Comics adaptation of The Empire Strikes Back from 1980.
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Pseudosymmetry, high copy number and twinning complicate the structure determination of Desulfovibrio desulfuricans (ATCC 29577) flavodoxin
Guelker, Megan
Stagg, Loren
Wittung-Stafshede, Pernilla
Shamoo, Yousif
Acta Crystallographica Section D: Biological Crystallography
2009;65(Pt 6):523-534.
Complications to molecular replacement resulting from a poor starting search model, pseudosymmetry, twinning and a high copy number in the asymmetric unit made the determination of the structure of D. desulfuricans (ATCC 29577) flavodoxin in two crystal forms challenging.
The crystal structure of oxidized flavodoxin from Desulfovibrio desulfuricans (ATCC 29577) was determined by molecular replacement in two crystal forms, P3121 and P43, at 2.5 and 2.0 Å resolution, respectively. Structure determination in space group P3121 was challenging owing to the presence of pseudo-translational symmetry and a high copy number in the asymmetric unit (8). Initial phasing attempts in space group P3121 by molecular replacement using a poor search model (46% identity) and multi-wavelength anomalous dispersion were unsuccessful. It was necessary to solve the structure in a second crystal form, space group P43, which was characterized by almost perfect twinning, in order to obtain a suitable search model for molecular replacement. This search model with complementary approaches to molecular replacement utilizing the pseudo-translational symmetry operators determined by analysis of the native Patterson map facilitated the selection and manual placement of molecules to generate an initial solution in the P3121 crystal form. During the early stages of refinement, application of the appropriate twin law, (−h, −k, l), was required to converge to reasonable R-factor values despite the fact that in the final analysis the data were untwinned and the twin law could subsequently be removed. The approaches used in structure determination and refinement may be applicable to other crystal structures characterized by these complicating factors. The refined model shows flexibility of the flavin mononucleotide coordinating loops indicated by the isolation of two loop conformations and provides a starting point for the elucidation of the mechanism used for protein-partner recognition.
flavodoxins; pseudosymmetry; twinning; high copy number; molecular replacement
Related Articles
Crystallization and preliminary crystallographic analysis of mouse peroxiredoxin II with significant pseudosymmetry
Ora, Ari
Oksanen, Esko
Kajander, Tommi
Goldman, Adrian
Butcher, Sarah J.
Acta Crystallographica Section F
2010;66(Pt 3):357-360.
Mouse peroxiredoxin II was crystallized in an orthorhombic space group and native X-ray diffraction data were collected.
Peroxiredoxin II was cloned from mouse B cells into pCold 1 expression vector and produced as a His-tagged recombinant protein in Escherichia coli. A ring form was isolated by gel filtration. A crystal obtained by the sitting-drop vapour-diffusion method diffracted to 1.77 Å resolution at 100 K. The crystal belonged to space group P21212, with unit-cell parameters a = 117.4, b = 133.9, c = 139.1 Å. The asymmetric unit is expected to contain six dimers of peroxiredoxin II, with a corresponding solvent content of 39.3%. Peaks in the native Patterson function together with pseudo-systematic absences suggested that the crystals suffered from severe translational pseudosymmetry.
peroxiredoxins; pseudosymmetry
Related Articles
Monoclinic crystal form of Aspergillus niger α-amylase in complex with maltose at 1.8 Å resolution
Vujičić-Žagar, A.
Dijkstra, B. W.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2006;62(Pt 8):716-721.
Two new crystal structures of A. niger α-amylase are reported, one of which reveals two hitherto unobserved maltose-binding sites.
Aspergillus niger α-amylase catalyses the hydrolysis of α-1,4-glucosidic bonds in starch. It shows 100% sequence identity to the A. oryzae homologue (also called TAKA-amylase), three crystal structures of which have been published to date. Two of them belong to the orthorhombic space group P212121 with one molecule per asymmetric unit and one belongs to the monoclinic space group P21 with three molecules per asymmetric unit. Here, the purification, crystallization and structure determination of A. niger α-amylase crystallized in the monoclinic space group P21 with two molecules per asymmetric unit in complex with maltose at 1.8 Å resolution is reported. Furthermore, a novel 1.6 Å resolution orthorhombic crystal form (space group P21212) of the native enzyme is presented. Four maltose molecules are observed in the maltose–α-amylase complex. Three of these occupy active-site subsites −2 and −1, +1 and +2 and the hitherto unobserved subsites +4 (Asp233, Gly234) and +5 (Asp235). The fourth maltose molecule binds at the distant binding sites d1 (Tyr382) and d2 (Trp385), also previously unobserved. Furthermore, it is shown that the active-site groove permits different binding modes of sugar units at subsites +1 and +2. This flexibility of the active-site cleft close to the catalytic centre might be needed for a productive binding of substrate chains and/or release of products.
α-amylase; Aspergillus niger; maltose; Aspergillus oryzae; TAKA-amylase
Related Articles
Crystallization and initial X-ray diffraction studies of the flavoenzyme NAD(P)H:(acceptor) oxidoreductase (FerB) from the soil bacterium Paracoccus denitrificans
Klumpler, Tomáš
Sedláček, Vojtěch
Marek, Jaromír
Wimmerová, Michaela
Kučera, Igor
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2010;66(Pt 4):431-434.
The flavin-dependent enzyme FerB from P. denitrificans has been purified and both native and SeMet-substituted FerB have been crystallized. The two variants crystallized in two different crystallographic forms belonging to the monoclinic space group P21 and the orthorhombic space group P21212, respectively. X-ray diffraction data were collected to 1.75 Å resolution for both forms.
The flavin-dependent enzyme FerB from Paracoccus denitrificans reduces a broad range of compounds, including ferric complexes, chromate and most notably quinones, at the expense of the reduced nicotinamide adenine dinucleotide cofactors NADH or NADPH. Recombinant unmodified and SeMet-substituted FerB were crystallized under similar conditions by the hanging-drop vapour-diffusion method with microseeding using PEG 4000 as the precipitant. FerB crystallized in several different crystal forms, some of which diffracted to approximately 1.8 Å resolution. The crystals of native FerB belonged to space group P21, with unit-cell parameters a = 61.6, b = 110.1, c = 65.2 Å, β = 118.2° and four protein molecules in the asymmetric unit, whilst the SeMet-substituted form crystallized in space group P21212, with unit-cell parameters a = 61.2, b = 89.2, c = 71.5 Å and two protein molecules in the asymmetric unit. Structure determination by the three-wavelength MAD/MRSAD method is now in progress.
flavoenzymes; quinone reductases; Paracoccus denitrificans
Related Articles
Imperfect pseudo-merohedral twinning in crystals of fungal fatty acid synthase
Jenni, Simon
Ban, Nenad
Acta Crystallographica Section D: Biological Crystallography
2009;65(Pt 2):101-111.
A case of imperfect pseudo-merohedral twinning in monoclinic crystals of fungal fatty acid synthase is discussed. A space-group transition during crystal dehydration resulted in a Moiré pattern-like interference of the twinned diffraction patterns.
The recent high-resolution structures of fungal fatty acid synthase (FAS) have provided new insights into the principles of fatty acid biosynthesis by large multifunctional enzymes. The crystallographic phase problem for the 2.6 MDa fungal FAS was initially solved to 5 Å resolution using two crystal forms from Thermomyces lanuginosus. Monoclinic crystals in space group P21 were obtained from orthorhombic crystals in space group P212121 by dehydration. Here, it is shown how this space-group transition induced imperfect pseudo-merohedral twinning in the monoclinic crystal, giving rise to a Moiré pattern-like interference of the two twin-related reciprocal lattices. The strategy for processing the twinned diffraction images and obtaining a quantitative analysis is presented. The twinning is also related to the packing of the molecules in the two crystal forms, which was derived from self-rotation function analysis and molecular-replacement solutions using a low-resolution electron microscopy map as a search model.
imperfect pseudo-merohedral twinning; fungal fatty acid synthase
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The pseudosymmetric structure of bis(pentane-1,5-diaminium) iodide tris(triiodide)
van Megen, Martin
Reiss, Guido J.
Acta Crystallographica Section E: Structure Reports Online
2012;68(Pt 5):o1331-o1332.
The asymmetric unit of the title compound, [H3N(CH2)5NH3]2I[I3]3 or 2C5H16N2 2+·3I3 −·I−, consists of two crystallographically independent pentane-1,5-diaminium dications and two triiodide anions in general positions besides two additional triiodide and two iodide anions located on twofold axes. The compound crystallizes in the centrosymmetric monoclinic space group P2/n. The structure refinement was handicapped by the pseudosymmetry (pseudo-centering) of the structure and by twinning. The crystal structure is composed of two alternate layers, which differ in their arrangement of the pentane-1,5-diaminium dications and the iodide/triiodide anions and which are connected via weak to medium–strong N—H⋯I hydrogen bonds, constructing a complex hydrogen-bonded network.
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Surprises and pitfalls arising from (pseudo)symmetry
Zwart, Peter H.
Grosse-Kunstleve, Ralf W.
Lebedev, Andrey A.
Murshudov, Garib N.
Adams, Paul D.
Acta Crystallographica Section D: Biological Crystallography
2007;64(Pt 1):99-107.
The presence of pseudosymmetry can cause problems in structure determination and refinement. The relevant background and representative examples are presented.
It is not uncommon for protein crystals to crystallize with more than a single molecule per asymmetric unit. When more than a single molecule is present in the asymmetric unit, various pathological situations such as twinning, modulated crystals and pseudo translational or rotational symmetry can arise. The presence of pseudosymmetry can lead to uncertainties about the correct space group, especially in the presence of twinning. The background to certain common pathologies is presented and a new notation for space groups in unusual settings is introduced. The main concepts are illustrated with several examples from the literature and the Protein Data Bank.
pathology; twinning; pseudosymmetry
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Pseudo-merohedral twinning and noncrystallographic symmetry in orthorhombic crystals of SIVmac239 Nef core domain bound to different-length TCRζ fragments
Kim, Walter M.
Sigalov, Alexander B.
Stern, Lawrence J.
Acta Crystallographica Section D: Biological Crystallography
2010;66(Pt 2):163-175.
P212121 crystals of SIV Nef core domain bound to a peptide fragment of the T-cell receptor ζ subunit exhibited noncrystallographic symmetry and nearly perfect pseudo-merohedral twinning simulating tetragonal symmetry. For a different peptide fragment, nontwinned tetragonal crystals were observed but diffracted to lower resolution. The structure was determined after assignment of the top molecular-replacement solutions to various twin or NCS domains followed by refinement under the appropriate twin law.
HIV/SIV Nef mediates many cellular processes through interactions with various cytoplasmic and membrane-associated host proteins, including the signalling ζ subunit of the T-cell receptor (TCRζ). Here, the crystallization strategy, methods and refinement procedures used to solve the structures of the core domain of the SIVmac239 isolate of Nef (Nefcore) in complex with two different TCRζ fragments are described. The structure of SIVmac239 Nefcore bound to the longer TCRζ polypeptide (Leu51–Asp93) was determined to 3.7 Å resolution (R work = 28.7%) in the tetragonal space group P43212. The structure of SIVmac239 Nefcore in complex with the shorter TCRζ polypeptide (Ala63–Arg80) was determined to 2.05 Å resolution (R work = 17.0%), but only after the detection of nearly perfect pseudo-merohedral crystal twinning and proper assignment of the orthorhombic space group P212121. The reduction in crystal space-group symmetry induced by the truncated TCRζ polypeptide appears to be caused by the rearrangement of crystal-contact hydrogen-bonding networks and the substitution of crystallographic symmetry operations by similar noncrystallographic symmetry (NCS) operations. The combination of NCS rotations that were nearly parallel to the twin operation (k, h, −l) and a and b unit-cell parameters that were nearly identical predisposed the P212121 crystal form to pseudo-merohedral twinning.
pseudo-merohedral twinning; noncrystallographic symmetry; pseudosymmetry; human immunodeficiency virus; Nef; T-cell receptor
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Crystallization, data collection and phasing of two digestive lysozymes from Musca domestica
Marana, S. R.
Cançado, F. C.
Valério, A. A.
Ferreira, C.
Terra, W. R.
Barbosa, J. A. R. G.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2006;62(Pt 8):750-752.
The digestive lysozymes 1 and 2 from M. domestica were crystallized by vapour diffusion. The crystallographic data were processed to a maximum resolution of 1.9 Å in both cases.
Lysozymes are mostly known for their defensive role against bacteria, but in several animals lysozymes have a digestive function. Here, the initial crystallographic characterization of two digestive lysozymes from Musca domestica are presented. The proteins were crystallized using the sitting-drop vapour-diffusion method in the presence of ammonium sulfate or PEG/2-propanol as the precipitant. X-ray diffraction data were collected to a maximum resolution of 1.9 Å using synchrotron radiation. The lysozyme 1 and 2 crystals belong to the monoclinic space group P21 (unit-cell parameters a = 36.52, b = 79.44, c = 45.20 Å, β = 102.97°) and the orthorhombic space group P21212 (unit-cell parameters a = 73.90, b = 96.40, c = 33.27 Å), respectively. The crystal structures were solved by molecular replacement and structure refinement is in progress.
digestive lysozymes; Musca domestica
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A novel monoclinic phase of impurity-doped CaGa2S4 as a phosphor with high emission intensity
Suzuki, Akihiro
Takizawa, Takeo
Hidaka, Chiharu
Shigetaka, Nomura
Kitajima, Ittetsu
Acta Crystallographica Section E: Structure Reports Online
2012;68(Pt 6):i42.
In the solid-state synthesis of impurity-doped CaGa2S4, calcium tetrathiodigallate(III), a novel phosphor material (denominated as the X-phase), with monoclinic symmetry in the space group P21/a, has been discovered. Its emission intensity is higher than that of the known orthorhombic polymorph of CaGa2S4 crystallizing in the space group Fddd. The asymmetric unit of the monoclinic phase consists of two Ca, four Ga and eight S sites. Each of the Ca and Ga atoms is surrounded by seven and four sulfide ions, respectively, thereby sharing each of the sulfur sites with the nearest neighbours. In contrast, the corresponding sites in the orthorhombic phase are surrounded by eight and four S atoms, respectively. The photoluminescence peaks from Mn2+ and Ce3+ in the doped X-phase, both of which are supposed to replace Ca2+ ions, have been observed to shift towards the high energy side in comparison with those in the orthorhombic phase. This suggests that the crystal field around the Mn2+ and Ce3+ ions in the X-phase is weaker than that in the orthorhombic phase.
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Purification, crystallization and preliminary X-ray analysis of Enterococcus casseliflavus aminoglycoside-2′′-phosphotransferase-IVa
Toth, Marta
Vakulenko, Sergei
Smith, Clyde A.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2009;66(Pt 1):81-84.
Aminoglycoside-2′′-phosphotransferase-IVa [APH(2′′)-IVa] is an enzyme that is responsible for high-level gentamicin resistance in E. casseliflavus isolates. Three different crystals of wild-type substrate-free APH(2′′)-IVa have been prepared and preliminary X-ray diffraction experiments have been undertaken on all three crystal forms.
The deactivation of aminoglycoside antibiotics by chemical modification is one of the major sources of bacterial resistance to this family of therapeutic compounds, which includes the clinically relevant drugs streptomycin, kanamycin and gentamicin. The aminoglycoside phosphotransferases (APHs) form one such family of enzymes responsible for this resistance. The gene encoding one of these enzymes, aminoglycoside-2′′-phosphotransferase-IVa [APH(2′′)-IVa] from Enterococcus casseliflavus, has been cloned and the protein (comprising 306 amino-acid residues) has been expressed in Escherichia coli and purified. The enzyme was crystallized in three substrate-free forms. Two of the crystal forms belonged to the orthorhombic space group P212121 with similar unit-cell parameters, although one of the crystal forms had a unit-cell volume that was approximately 13% smaller than the other and a very low solvent content of around 38%. The third crystal form belonged to the monoclinic space group P21 and preliminary X-ray diffraction analysis was consistent with the presence of two molecules in the asymmetric unit. The orthorhombic crystal forms of apo APH(2′′)-IVa both diffracted to 2.2 Å resolution and the monoclinic crystal form diffracted to 2.4 Å resolution; synchrotron diffraction data were collected from these crystals at SSRL (Stanford, California, USA). Structure determination by molecular replacement using the structure of the related enzyme APH(2′′)-IIa is proceeding.
aminoglycoside-2′′-phosphotransferase-IVa; Enterococcus casseliflavus; antibiotic resistance
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Pseudomerohedrally twinned monoclinic structure of unfolded 'free' nonactin: comparative analysis of its large conformational change upon encapsulation of alkali metal ions
Guzei, Ilia A.
Wang, Cheng
Zhan, Yulian
Dolomanov, Oleg V.
Cheng, Yi-Qiang
Acta Crystallographica Section C: Crystal Structure Communications
2009;65(Pt 10):o521-o524.
The title compound, C40H64O12, crystallizes in a pseudomerohedrally twinned primitive monoclinic cell with similar contributions of the two twin components. There are two symmetry-independent half-molecules of nonactin in the asymmetric unit. Each molecule has a pseudo-S 4 symmetry and resides on a crystallographic twofold axis; the axes pass through the molecular center of mass and are perpendicular to the plane of the macrocycle. The literature description of the room-temperature structure of nonactin as an order–disorder structure in an orthorhombic unit cell is corrected. We report a low-temperature high-precision ordered structure of 'free' nonactin that allowed for the first time precise determination of its bond distances and angles. It possesses an unfolded and more planar geometry than its complexes with encapsulated Na+, K+, Cs+, Ca2+ or NH4 + cations that exhibit more isometric overall conformations.
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Crystallographic characterization of two novel crystal forms of human insulin induced by chaotropic agents and a shift in pH
Norrman, Mathias
Schluckebier, Gerd
BMC Structural Biology
Insulin is a therapeutic protein that is widely used for the treatment of diabetes. Its biological function was discovered more than 80 years ago and it has since then been characterized extensively. Crystallization of the insulin molecule has always been a key activity since the protein is often administered by subcutaneous injections of crystalline insulin formulations. Over the years, insulin has been crystallized and characterized in a number of crystal systems.
Interestingly, we have now discovered two new crystal forms of human insulin. The crystals were obtained when the two chaotropic agents, urea and thiocyanate were present in the crystallization experiments, and their structures were determined by X-ray crystallography. The crystals belong to the orthorhombic and monoclinic crystal systems, with space groups C2221 and C2 respectively. The orthorhombic crystals were obtained at pH 6.5 and contained three insulin hexamers in R6 conformation in the asymmetric unit whilst the monoclinic C2 crystals were obtained at pH 7.0 and contained one R6 hexamer in the asymmetric unit. Common for the two new crystals is a hexamer-hexamer interaction that has not been found in any of the previous crystal forms of insulin. The contacts involve a tight glutamate-glutamate interaction with a distance of 2.3 Å between groups. The short distance suggests a low barrier hydrogen bond. In addition, two tyrosine-tyrosine interactions occupying a known phenol binding pocket contribute to the stabilization of the contacts. Within the crystals, distinct binding sites for urea were found, adding further to the discussion on the role of urea in protein denaturation.
The change in space group from C2221 to C2 was primarily caused by an increase in pH. The fewer number of hexamer-hexamer interactions comprising the short hydrogen bond in the C2 space group suggest that pH is the driving force. In addition, the distance between the two glutamates increases from 2.32 Å in the C2221 crystals to 2.4 Å in the C2 crystals. However, in both cases the low barrier hydrogen bond and the tyrosine-tyrosine interaction should contribute to the stability of the crystals which is crucial when used in pharmaceutical formulations.
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Cloning, expression, purification, crystallization and preliminary X-ray analysis of the human RuvBL1–RuvBL2 complex
Gorynia, Sabine
Matias, Pedro M.
Bandeiras, Tiago M.
Donner, Peter
Carrondo, Maria Arménia
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2008;64(Pt 9):840-846.
A truncated variant of the human RuvBL1–RuvBL2 complex was cloned, expressed, purified and crystallised. Synchrotron diffraction data to 4 Å resolution were used to carry out a preliminary crystallographic analysis of the complex.
The complex of RuvBL1 and its homologue RuvBL2, two evolutionarily highly conserved eukaryotic proteins belonging to the AAA+ (ATPase associated with diverse cellular activities) family of ATPases, was co-expressed in Escherichia coli. For crystallization purposes, the flexible domains II of RuvBL1 and RuvBL2 were truncated. The truncated RuvBL1–RuvBL2 complex was crystallized using the hanging-drop vapour-diffusion method at 293 K. The crystals were hexagonal-shaped plates and belonged to either the orthorhombic space group C2221, with unit-cell parameters a = 111.4, b = 188.0, c = 243.4 Å and six monomers in the asymmetric unit, or the monoclinic space group P21, with unit-cell parameters a = 109.2, b = 243.4, c = 109.3 Å, β = 118.7° and 12 monomers in the asymmetric unit. The crystal structure could be solved by molecular replacement in both possible space groups and the solutions obtained showed that the complex forms a dodecamer.
RuvBL1; RuvBL2; ATPases
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Crystallization and preliminary X-ray crystallographic analysis of a blue-light-absorbing proteorhodopsin
Wang, Ning
Wang, Meitian
Gao, Yanyan
Ran, Tingting
Lan, Yanli
Wang, Jian
Xu, Langlai
Wang, Weiwu
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2012;68(Pt 3):281-283.
A purified blue-light-absorbing proteorhodopsin D97N mutant protein (BPR_D97N) has been crystallized using the vapour-diffusion method.
Proteorhodopsins (PRs), seven-transmembrane chromoproteins with retinal as a chromophore, are light-driven proton pumps. To elucidate the light-driven proton-pumping mechanism of PRs, a pET28a vector containing the blue-light-absorbing proteorhodopsin (BPR) gene was constructed and the protein was overexpressed in Escherichia coli. The protein was purified by immobilized metal-ion affinity chromatography (IMAC). The purified BPR D97N mutant protein (BPR_D97N) was crystallized using the vapour-diffusion method. Preliminary X-ray diffraction data analysis showed that the crystal belonged to the orthorhombic space group P21212, with unit-cell parameters a = 161.6, b = 168.6, c = 64.7 Å. A complete data set was collected to 3.3 Å resolution using synchrotron radiation on beamline X06 of the Swiss Light Source (SLS). Molecular replacement was unsuccessful. To solve the structure of BPR_D97N by experimental phasing, selenomethionine-substituted protein crystals were prepared. These crystals diffracted to 3.0 Å resolution and a complete data set was collected on beamline BL17U of the Shanghai Synchrotron Radiation Facility (SSRF). Heavy-atom substructure determination and phasing by SAD clearly showed that the crystal contained five molecules in the asymmetric unit, with a V M of 3.26 Å3 Da−1 and a solvent content of 62.3%.
proteorhodopsins; BPR
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Structure determination of enterovirus 71
Plevka, Pavel
Perera, Rushika
Cardosa, Jane
Kuhn, Richard J.
Rossmann, Michael G.
Acta Crystallographica Section D: Biological Crystallography
2012;68(Pt 9):1217-1222.
Determination of the orientation of the enterovirus 71 virions in the crystal required the calculation of a locked rotation function that included only icosahedral threefold and fivefold symmetry axes. Otherwise, misleading high rotation-function values were produced by accidental alignment of icosahedral and crystallographic twofold axes.
Enterovirus 71 is a picornavirus that causes hand, foot and mouth disease but may induce fatal neurological illness in infants and young children. Enterovirus 71 crystallized in a body-centered orthorhombic space group with two particles in general orientations in the crystallographic asymmetric unit. Determination of the particle orientations required that the locked rotation function excluded the twofold symmetry axes from the set of icosahedral symmetry operators. This avoided the occurrence of misleading high rotation-function values produced by the alignment of icosahedral and crystallographic twofold axes. Once the orientations and positions of the particles had been established, the structure was solved by molecular replacement and phase extension.
enterovirus 71; viruses; rotation function; molecular replacement
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Crystallization and preliminary X-ray crystallographic analysis of the oxysterol-binding protein Osh3 from Saccharomyces cerevisiae
Tong, Junsen
Yang, Huiseon
Ha, Subin
Lee, Youngjin
Eom, Soo Hyun
Im, Young Jun
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2012;68(Pt 12):1498-1502.
The PH domain and ORD of the oxysterol-binding protein Osh3 from S. cerevisae were crystallized and X-ray diffraction data were collected.
Oxysterol-binding protein (OSBP) related proteins (ORPs) are conserved from yeast to humans and are implicated in regulation of sterol homeostasis and in signal transduction pathways. Osh3 of Saccharomyces cerevisiae is a pleckstrin-homology (PH) domain-containing ORP member that regulates phosphoinositide metabolism at endoplasmic reticulum–plasma membrane contact sites. The N-terminal PH domain of Osh3 was purified and crystallized as a lysozyme fusion and the resulting crystal diffracted to 2.3 Å resolution. The crystal belonged to the monoclinic space group C2, with unit-cell parameters a = 98.03, b = 91.31, c = 84.13 Å, β = 81.41°. With two molecules in the asymmetric unit, the Matthews coefficient was 3.13 Å3 Da−1. Initial attempts to solve the structure by molecular-replacement techniques using T4 lysozyme as a search model were successful. The C-terminal OSBP-related domain (OBD) of Osh3 was crystallized by the vapour-diffusion method and the resulting crystal diffracted to 1.5 Å resolution. The crystal was orthorhombic, belonging to space group P212121, with unit-cell parameters a = 41.57, b = 87.52, c = 100.58 Å. With one molecule in the asymmetric unit, the Matthews coefficient was 2.01 Å3 Da−1. Initial attempts to solve the structure by the single-wavelength anomalous dispersion technique using bromine were successful.
oxysterol-binding protein; Osh3; Saccharomyces cerevisiae
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Expression, purification, crystallization and preliminary X-ray diffraction studies of phosphoglycerate mutase from Staphylococcus aureus NCTC8325
Roychowdhury, Amlan
Kundu, Anirban
Gujar, Akanksha
Bose, Madhuparna
Das, Amit Kumar
Acta Crystallographica. Section F, Structural Biology Communications
2013;70(Pt 1):53-56.
The cloning, overexpression, purification, crystallization and preliminary X-ray crystallographic analysis of phosphoglycerate mutase from S. aureus NCTC8325 are reported.
Phosphoglycerate mutase (PGM) is a key enzyme in carbohydrate metabolism. It takes part in both glycolysis and gluconeogenesis. PGM from pathogenic Staphylococcus aureus (NCTC8325) was cloned in pQE30 expression vector overexpressed in Escherichia coli M15 (pREP4) cells and purified to homogeneity. The protein was crystallized from two different conditions, (i) 0.1 M HEPES pH 7.5, 20%(w/v) polyethylene glycol 10 000 and (ii) 0.2 M NaCl, 0.1 M bis-tris pH 6.5, 25%(w/v) polyethylene glycol 3350, at 25°C by the sitting-drop vapour-diffusion method. Crystals grown at pH 7.5 diffracted to 2.5 Å resolution and belonged to the orthorhombic space group P21212, with unit-cell parameters a = 77.0, b = 86.11, c = 94.07 Å. Crystals from the second condition at pH 6.5 diffracted to 2.00 Å resolution. These crystals belonged to the orthorhombic space group P212121, with unit-cell parameters a = 73.21, b = 81.75, c = 89.18 Å. X-ray diffraction data have been collected and processed to the maximum resolution to determine the structure of PGM. The structure has been solved by molecular replacement and structure refinement is now in progress.
phosphoglycerate mutase; Staphylococcus aureus NCTC8325
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Structure of the orthorhombic form of human inosine triphosphate pyrophosphatase
Porta, Jason
Kolar, Carol
Kozmin, Stanislav G.
Pavlov, Youri I.
Borgstahl, Gloria E. O.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2006;62(Pt 11):1076-1081.
X-ray crystallographic analysis of human inosine triphosphate pyrophosphohydrolase provided the secondary structure and active-site structure at 1.6 Å resolution in an orthorhombic crystal form. The structure gives a framework for future structure–function studies employing site-directed mutagenesis and for the identification of substrate/product-binding sites.
The structure of human inosine triphosphate pyrophosphohydrolase (ITPA) has been determined using diffraction data to 1.6 Å resolution. ITPA contributes to the accurate replication of DNA by cleansing cellular dNTP pools of mutagenic nucleotide purine analogs such as dITP or dXTP. A similar high-resolution unpublished structure has been deposited in the Protein Data Bank from a monoclinic and pseudo-merohedrally twinned crystal. Here, cocrystallization of ITPA with a molar ratio of XTP appears to have improved the crystals by eliminating twinning and resulted in an orthorhombic space group. However, there was no evidence for bound XTP in the structure. Comparison with substrate-bound NTPase from a thermophilic organism predicts the movement of residues within helix α1, the loop before α6 and helix α7 to cap off the active site when substrate is bound.
inosine triphosphate pyrophosphohydrolase
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Superoxide reductase from Nanoarchaeum equitans: expression, purification, crystallization and preliminary X-ray crystallographic analysis
Pinho, Filipa G.
Pinto, Ana F.
Pinto, Liliana C.
Huber, Harald
Romão, Célia V.
Teixeira, Miguel
Matias, Pedro M.
Bandeiras, Tiago M.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2011;67(Pt 5):591-595.
Diffraction-quality crystals of N. equitans neelaredoxin have been produced. The expression, purification and crystallization of the protein and preliminary X-ray crystallographic analysis of the crystals are reported.
Superoxide reductases (SORs) are the most recent oxygen-detoxification system to be identified in anaerobic and microaerobic bacteria and archaea. SORs are metalloproteins that are characterized by their possession of a catalytic nonhaem iron centre in the ferrous form coordinated by four histidine ligands and one cysteine ligand. Ignicoccus hospitalis, a hyperthermophilic crenarchaeon, is the only organism known to date to serve as a host for Nanoarchaeum equitans, a nanosized hyperthermophilic archaeon isolated from a submarine hot vent which completely depends on the presence of and contact with I. hospitalis cells for growth to occur. Similarly to I. hospitalis, N. equitans has a neelaredoxin (a 1Fe-type SOR) that keeps toxic oxygen species under control, catalysing the one-electron reduction of superoxide to hydrogen peroxide. Blue crystals of recombinant N. equitans SOR in the oxidized form (12.7 kDa, 109 residues) were obtained using polyethylene glycol (PEG 2000 MME) as precipitant. These crystals diffracted to 1.9 Å resolution at 100 K and belonged to the orthorhombic space group P212121, with unit-cell parameters a = 51.88, b = 82.01, c = 91.30 Å. Cell-content analysis suggested the presence of four monomers in the asymmetric unit. The Matthews coefficient (V M) was determined to be 1.9 Å3 Da−1, corresponding to an estimated solvent content of 36%. Self-rotation function and native Patterson calculations suggested a tetramer with 222 point-group symmetry, similar to other 1Fe-SORs. The three-dimensional structure will be determined by the molecular-replacement method.
superoxide reductases; Nanoarchaeum equitans; oxidative stress; neelaredoxins
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Expression, purification, crystallization and structure determination of two glutathione S-transferase-like proteins from Shewanella oneidensis
Remmerie, Bert
Vandenbroucke, Karen
De Smet, Lina
Carpentier, Wesley
De Vos, Dirk
Stout, Jan
Van Beeumen, Jozef
Savvides, Savvas N.
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2008;64(Pt 6):548-553.
The production and purification of recombinant SoGST3 and SoGST6, two GST-like proteins from S. oneidensis, are reported and preliminary crystallographic studies of crystals of the recombinant enzymes are presented.
Genome analysis of Shewanella oneidensis, a Gram-negative bacterium with an unusual repertoire of respiratory and redox capabilities, revealed the presence of six glutathione S-transferase-like genes (sogst1–sogst6). Glutathione S-transferases (GSTs; EC 22.214.171.124) are found in all kingdoms of life and are involved in phase II detoxification processes by catalyzing the nucleophilic attack of reduced glutathione on diverse electrophilic substrates, thereby decreasing their reactivity. Structure–function studies of prokaryotic GST-like proteins are surprisingly underrepresented in the scientific literature when compared with eukaryotic GSTs. Here, the production and purification of recombinant SoGST3 (SO_1576) and SoGST6 (SO_4697), two of the six GST-like proteins in S. oneidensis, are reported and preliminary crystallographic studies of crystals of the recombinant enzymes are presented. SoGST3 was crystallized in two different crystal forms in the presence of GSH and DTT that diffracted to high resolution: a primitive trigonal form in space group P31 that exhibited merohedral twinning with a high twin fraction and a primitive monoclinic form in space group P21. SoGST6 yielded primitive orthorhombic crystals in space group P212121 from which diffraction data could be collected to medium resolution after application of cryo-annealing protocols. Crystal structures of both SoGST3 and SoGST6 have been determined based on marginal search models by maximum-likelihood molecular replacement as implemented in the program Phaser.
glutathione S-transferases; Shewanella oneidensis
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An orthorhombic polymorph of pyrazino[2,3-f][1,10]phenanthroline-2,3-dicarbonitrile
Yang, Wei
Qiu, Qi-Ming
Zhou, Li-Li
Jin, Qiong-Hua
Zhang, Cun-Lin
Acta Crystallographica Section E: Structure Reports Online
2011;67(Pt 12):o3250-o3251.
The title compound, C16H6N6, is a polymorph of the previously reported structure [Kozlov & Goldberg (2008 ▶). Acta Cryst. C64, o498–o501]. Unlike the previously reported monoclinic polymorph (space group P21/c, Z = 8), the title compound reveals orthorhombic symmetry (space group Pnma, Z = 4). The molecule shows crystallographic mirror symmetry, while the previously reported structure exhibits two independent molecules per asymmetric unit. In the title compound, adjacent molecules are essentially parallel along the c axis and tend to be vertical along the b axis with dihedral angles of 72.02 (6)°. However, in the reported polymorph, the entire crystal structure shows an antiparallel arrangement of adjacent columns related by inversion centers and the two independent molecules are nearly parallel with a dihedral angle of 2.48 (6)°.
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From small structural modifications to adjustment of structurally dependent properties: 1-methyl-3,5-bis[(E)-2-thienylidene]-4-piperidone and 3,5-bis[(E)-5-bromo-2-thienylidene]-1-methyl-4-piperidone
Tongwa, Paul
Kinnibrugh, Tiffany L.
Kicchaiahgari, Geetha R.
Khrustalev, Victor N.
Timofeeva, Tatiana V.
Acta Crystallographica Section C: Crystal Structure Communications
2009;65(Pt 4):o155-o159.
The molecules of the title compounds, C16H15NOS2, (I), and C16H13Br2NOS2, (II), are E,E-isomers and consist of an extensive conjugated system, which determines their molecular geometries. Compound (I) crystallizes in the monoclinic space group P21/c. It has one thiophene ring disordered over two positions, with a minor component contribution of 0.100 (3). Compound (II) crystallizes in the noncentrosymmetric orthorhombic space group Pca21 with two independent molecules in the unit cell. These molecules are related by a noncrystallographic pseudo-inversion center and possess very similar geometries. The crystal packings of (I) and (II) have a topologically common structural motif, viz. stacks along the b axis, in which the molecules are bound by weak C—H⋯O hydrogen bonds. The noncentrosymmetric packing of (II) is governed by attractive intermolecular Br⋯Br and Br⋯N interactions, which are also responsible for the very high density of (II) (1.861 Mg m−3).
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Monohalogenated ferrocenes C5H5FeC5H4 X (X = Cl, Br and I) and a second polymorph of C5H5FeC5H4I
Romanov, Alexander S.
Mulroy, Joseph M.
Khrustalev, Victor N.
Antipin, Mikhail Yu.
Timofeeva, Tatiana V.
Acta Crystallographica Section C: Crystal Structure Communications
2009;65(Pt 11):m426-m430.
The structures of the three title monosubstituted ferrocenes, namely 1-chloroferrocene, [Fe(C5H5)(C5H4Cl)], (I), 1-bromoferrocene, [Fe(C5H5)(C5H4Br)], (II), and 1-iodoferrocene, [Fe(C5H5)(C5H4I)], (III), were determined at 100 K. The chloro- and bromoferrocenes are isomorphous crystals. The new triclinic polymorph [space group P , Z = 4, T = 100 K, V = 943.8 (4) Å3] of iodoferrocene, (III), and the previously reported monoclinic polymorph of (III) [Laus, Wurst & Schottenberger (2005 ▶). Z. Kristallogr. New Cryst. Struct. 220, 229–230; space group Pc, Z = 4, T = 100 K, V = 924.9 Å3] were obtained by crystallization from ethanolic solutions at 253 and 303 K, respectively. All four phases contain two independent molecules in the unit cell. The relative orientations of the cyclopentadienyl (Cp) rings are eclipsed and staggered in the independent molecules of (I) and (II), while (III) demonstrates only an eclipsed conformation. The triclinic and monoclinic polymorphs of (III) contain nonbonded intermolecular I⋯I contacts, causing different packing modes. In the triclinic form of (III), the molecules are arranged in zigzag tetramers, while in the monoclinic form the molecules are arranged in zigzag chains along the a axis. Crystallographic data for (III), along with the computed lattice energies of the two polymorphs, suggest that the monoclinic form is more stable.
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Expression, purification, crystallization and preliminary X-ray diffraction analysis of the apo form of InsP5 2-K from Arabidopsis thaliana
Baños-Sanz, Jose Ignacio
Sanz-Aparicio, Julia
Brearley, Charles A.
González, Beatriz
Acta Crystallographica Section F: Structural Biology and Crystallization Communications
2012;68(Pt 6):701-704.
A mutated version of InsP5 2-K allows the production of crystals of the apo form and structure determination using X-ray crystallography.
Inositol 1,3,4,5,6-pentakisphosphate 2-kinase (IP5 2-K) is a key enzyme that catalyzes the synthesis of phytic acid (IP6) from inositol 1,3,4,5,6-pentakisphosphate (IP5) and ATP. The first structure of IP5 2-K, that from Arabidopsis thaliana, has been solved previously; it only crystallized in the presence of inositol, either the substrate IP5 or the product IP6, and failed to crystallize in its free state (without inositol). Based on structural analysis, a point mutation of IP5 2-K (W129A) has been produced in order to overcome this limitation and obtain information about protein conformational changes upon substrate binding. Here, the production and crystallization of W129A IP5 2-K in its free state and with bound nucleotide is described. These crystals differed from the native crystals and belonged to the orthorhombic space group P21212, with unit-cell parameters a = 66.00, b = 68.23, c = 105.80 Å and a = 63.06, b = 71.80, c = 100.23 Å, respectively. The crystals diffracted to resolutions of 2.22 Å (apo) and 2.05 Å (nucleotide bound) using synchrotron radiation and contained one molecule per asymmetric unit. The structures have been determined using the molecular-replacement method and refinement is being undertaken.
inositol kinases; inositol phosphate; phytic acid; IP6; IP5 2-K
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"I'm very pleased but I was not too worried about the result as I just wanted the lads to get back to playing football before next week's Premier cup final."
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"God is not pleased with these mashed potato Christians. You know the kind that are found in the T.V. dinners. They are hot on the outside but ice cold on the inside. They look good but they are of no use and usually get thrown out!"
"This was the result the lads deserved. They've worked hard all season. We can now go on from this."
"The lads treated the game as a cup final and the way they played I cannot fault them. The result was better late than never and went a long way to getting the monkey off our back."
"And he said, Lay not thine hand upon the lad, neither do thou any thing unto him: for now I know that thou fearest God, seeing thou hast not withheld thy son, thine only son from me."
"He is back fit now, which I am pleased about. He is a powerful lad, who seems confident after a good run in the team, and quite right too."


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Supreme Court of New York, First Department
May 14, 2013
20-35 86th Street Realty, LLC, Plaintiff-Appellant,v.Tower Insurance Company of New York, Defendant-Respondent.
Razis & Ross, P.C., Astoria (George J. Razis of counsel), for appellant.
Mound Cotton Wollan & Greengrass, New York (Tania A. Gondiosa of counsel), for respondent.
Andrias, J.P., Saxe, Freedman, Román, JJ.
Order, Supreme Court, New York County (Debra A. James, J.), entered April 17, 2012, which granted defendant's motion to dismiss the complaint to the extent of staying the action pending the determination of the criminal case against the alleged arsonist, and denied plaintiff's motion to dismiss the second, third, fourth, fifth and sixth affirmative defenses, unanimously modified, on the law, to grant plaintiff's motion as to the second, third and sixth affirmative defenses, and to vacate the stay, and otherwise affirmed, without costs.
Plaintiff building owner seeks a judgment declaring that defendant has an obligation to defend and indemnify it in connection with a building fire that, inter alia, left several people dead or injured. Following a police and fire department investigation, an individual was arrested and indicted on charges of, inter alia, arson in the fourth degree and assault in the first and second degrees. Defendant disclaimed coverage under the liability insurance policy it issued to plaintiff on the ground that plaintiff's claims apparently involved bodily injury arising from an assault or battery committed by the accused arsonist and that the insurance does not apply to bodily injury arising from assault and/or battery or any act or omission in connection with any assault and/or battery. The motion court stayed this action pending determination of the criminal action on the ground that that determination is necessary to a determination of the applicability of the assault and battery exclusion to plaintiff's claims.
Civil assault and battery are intentional acts, and the assault offenses with which the accused arsonist is charged do not include the intent to harm a specific individual ( compare PJI 2d 3:2 [assault]; 3:3 [battery], with Penal Law 120.10[4] [assault in the first degree]; 120.05[6] [assault in the second degree]). Thus, assuming that the insurance policy exclusion is triggered by civil, rather than criminal, assault or battery, the critical inquiry is whether the accused arsonist, in allegedly causing the fire, intended to harm any occupant of the building. Although the determination of the criminal action is therefore not necessary to a determination of the application of the exclusion, the criminal trial may shed light on the accused arsonist's motives, including whether he intended to harm anyone inside the building. In any event, the criminal trial may enable defendant to obtain access to evidence and witnesses that will assist in determining whether the exclusion applies. Based on representations made at oral argument, the criminal trial has been concluded and, thus, the stay should be lifted.
In light of the foregoing, the motion court correctly denied plaintiff's motion to dismiss the affirmative defenses based on the assault and battery exclusion and the lack of bodily injury caused by an accident or occurrence. However, defendant did not refer in its disclaimer of coverage to a failure to comply with the policy terms or a failure to cooperate, and those grounds may not be asserted as affirmative defenses ( see Insurance Law § 3420[d]; Benjamin Shapiro Realty Co. v Agricultural Ins. Co., 287 A.D.2d 389 [1st Dept 2001]). Defendant's recourse for an insufficiently specific complaint was to move under CPLR 3024(a) for a more definite statement or to amend its answer as of right under CPLR 3025, within 20 days after service of the answer, or, after 20 days, move for leave to amend. Defendant may not override the statute by reserving a right to amend in its answer.
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August 17, 2010
MARCELLA DANIELS, PLAINTIFF,v.FREEDMAN ANSELMO LINDBERG & RAPPE LLC, ET AL., DEFENDANTS.
Marcella Daniels ("Daniels") initiated this lawsuit by filing a pro se Complaint against a number of defendants, seeking to invoking the Fair Debt Collection Practices Act and the Fair Credit Reporting Act as the basis for federal jurisdiction. Daniels also filled out and tendered two forms provided by this District Court's Clerk's Office: an In Forma Pauperis Application ("Application") and a Motion for Appointment of Counsel ("Motion"). This Court promptly issued a March 29, 2010 memorandum order that found (1) Daniels had failed to qualify for in forma pauperis treatment so far as nonpayment of the filing fee was concerned and (2) Daniels' Motion was deficient in terms of a failure to provide the information that our Court of Appeals teaches is an essential component of such a request.
Daniels then paid the filing fee and then moved on June 23 to have the United States Marshals Service serve the defendants. This Court denied that motion orally on the date of its presentment, explaining to Daniels the narrow statutory authorization for service by the Marshals Service.
Now Daniels has submitted a new Motion, listing four lawyers with whom she has spoken about representation and stating:
Some lawyers that I called fees was very high and at this time I'm not working and can't afford it.
For much the same reason that prompted denial of the Application, this Court finds that Daniels has not satisfactorily shown "poverty" (the relevant precondition called for by 28 U.S.C. §1915(e)). Accordingly the Motion is denied. Daniels, who has shown herself to be articulate and has exhibited a full grasp of her claims in her in-court appearances,*fn1 will continue to proceed pro se.
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Property Location With a stay at Nautical Faralya in Fethiye (Faralya), you'll be convenient to Butterfly Valley and Kabak Beach. This 4-star hotel is within the vicinity of Oludeniz Beach and Oludeniz National Park.Rooms Make yourself at home in one of the 13 air-conditioned rooms featuring minibars. Rooms have private furnished patios. Complimentary wireless Internet access is available to keep you connected. Private bathrooms with showers feature hair dryers and bathrobes.Rec, Spa, Premium Amenities Head down to the water and enjoy a day at the private beach, or take advantage of other recreational amenities, which include a seasonal outdoor pool.Dining Grab a bite to eat at the hotel's restaurant, where you can enjoy drinks at a bar and dine alfresco (weather permitting). Or stay in and take advantage of room service (during limited hours). Quench your thirst with your favorite drink at a bar/lounge. A complimentary full breakfast is served daily.Business, Other Amenities Featured amenities include a 24-hour front desk and coffee/tea in the lobby. A roundtrip airport shuttle is provided for a surcharge (available on request).
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A fantastic place with the best sunset in the world every nite super chill a very special place
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Tabiat ile iç içe, insan kalabalığı olmayan, otel içindeki her metrekarenin sizin konforunuza göre düZenlendiğini anladığınız, hizmette hızlı ve güleryüzlü müthiş bir butik otel
- ibeking
- 2013-08-22
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daha temiz olabilirdi.
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- 2013-10-04
- 0 likes


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Gender: Girl
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No Cook Divinity
Barbara Bennett
Easy recipe if you are a divinity fan. You can add other things to your taste. Enjoy!
Featured Pinch Tips Video
- 1
- package white frosting
- 1/3 c
- light corn syrup
- 2 Tbsp
- boiling water
- 1 tsp
- vanilla
- 3 3/4 c
- powdered sugar
- 6 Tbsp
- hot water
- 1 c
- chopped nuts
Directions Step-By-Step
1In a large mixing bowl, combine the frosting, corn syrup, boiling water and vanilla. Beat on high for 5 minutes. Gradually add the powdered sugar, hot water and nuts; mix well. Drop by teaspoons onto waxed paper.Let stand at room temperature overnight or until dry to touch.Store in airtight container.
About this Recipe
Course/Dish: Candies
Main Ingredient: Dairy
Regional Style: American
Other Tag: Quick & Easy


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Fabrication of a hydraulic fork lift
This mechanical engineering project is about designing a hydraulic fork lift. There is nothing new about this project. I am sure that similar projects are done in many colleges. Even ready-made material may be available about the whole project. Anyway I don't have it so let me share what I think is worthy of this project.
Fabrication of the whole forklift including the cab and engine will be expensive. It won't much add to your academics either as what you plan to build is already in the market with a lot of advanced features. So better take this as a design project with calculations of the power, capacity, mobility etc. You can also fabricate a forklift with carriage (the portion that houses lifting fork, hydraulic cylinder and piston)
As far … Read the rest


text: KIOS News The Douglas County Election Commission announces new options for voters By Deborah (Van Fleet) Newcombe • Aug 29, 2016 Douglas and Sarpy County voters are receiving three-part postcards in the mail which include all the key points about voting for the November 8th general election. Omaha Reads "All the Light We Cannot See" By Cheril Lee • Aug 29, 2016 All the Light We Cannot See by Anthony Doerr is this year's Omaha Reads selection. Residents who receive SNAP benefits can apply to replace food lost after power outage By Cheril Lee & Nebraska DHHS • Aug 26, 2016 After Tuesday's storm, many residents were left without power and saw their food spoil. Midland University leases west Omaha space for MBA program By Associated Press • Aug 26, 2016 OMAHA, Neb. (AP) — Midland University has signed a decade-long lease in west Omaha for its MBA program and other offerings. Nebraska Sen. Matt Williams of Gothenburg to run for speaker By Associated Press • Aug 26, 2016 LINCOLN, Neb. (AP) — A state senator from Gothenburg has joined the race to become the Nebraska Legislature's next speaker. Reception for the UNO Women's Archive Project set for tonight By Cheril Lee • Aug 26, 2016 The UNO Women's Archive Project will have a reception this evening starting at 7:00 at KANEKO. Utility says some people may lack power until this afternoon By Associated Press • Aug 25, 2016 OMAHA, Neb. (AP) Several thousand utility customers in eastern Nebraska could remain without power until Thursday afternoon because of the extent of damage thunderstorms caused. 3 arrested when 22 lbs. of pot, 1 lb. of meth found By Associated Press • Aug 25, 2016 OMAHA, Neb. (AP) â Three people are facing charges after Douglas County Sheriff's deputies found 22 pounds of marijuana and 1 pound of methamphetamine in two different traffic stops this week. UNMC receives grant that will allow pharmacies to help track patient vaccinations By Deborah (Van Fleet) Newcombe • Aug 25, 2016 With more and more people getting immunizations at their local pharmacies, having access to accurate records about who needs what can be challenging. US Mint designs 3 coins for Boys Town's 100th anniversary By Associated Press • Aug 24, 2016 BOYS TOWN, Neb. (AP) — Three U.S. coins will be released next year to celebrate the 100th anniversary of Boys Town. Storms knock out power, flood streets in eastern Nebraska By Associated Press • Aug 24, 2016 OMAHA, Neb. (AP) — Thunderstorms have lashed several parts of eastern Nebraska, leaving thousands of people without power and some stranded by street flooding and storm debris. Do Space holds a Startup Summit next month By Cheril Lee • Aug 24, 2016 Do Space is holding a Startup Summit on Thursday, September 22nd from 11:30 to 1. Lunch will be provided. 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(AP) — Groups that want to legalize casinos at Nebraska horse racing tracks are trying to decide what to do now that a $1.4 million petition drive has failed to place the issue on the ballot. ConAgra Foods will continue its support of Shine the Light on Hunger, Omaha Holiday Lights Festival By Cheril Lee • Aug 22, 2016 ConAgra's 2016 Shine the Light on Hunger Campaign as well as their involvement in the Holiday Lights Festival will continue. Traffic Incident Management a public/private partnership By Cheril Lee • Aug 22, 2016 Approximately 40 agencies and organizations, both private and public, are working together to help reduce the duration of traffic incidents and the congestion caused by them. You Drink, You Drive, You Lose By Cheril Lee • Aug 22, 2016 The Omaha Police Department is taking part in a nationwide campaign called, "You Drink, You Drive, You Lose." "FiFi" arrives in Omaha and welcomes visitors By Chris Cooke • Aug 19, 2016 Commemorative Air Force's (CAF) iconic Boeing B-29 Superfortress "FiFi" is in Omaha through Sunday as part of the AirPower History Tour. AV Sorensen Library Celebrates its 40th anniversary By Deborah (Van Fleet) Newcombe • Aug 19, 2016 The AV Sorensen Library and Recreation Center, at 4808 Cass Street, will have an open house tomorrow from 10 to noon to celebrate its 40th anniversary. Omaha Public Radio's Deborah Newcombe has more… Survey says economy remains weak in rural parts of 10 states By Associated Press • Aug 19, 2016 OMAHA, Neb. (AP) — A monthly survey of bankers suggests the economy remains weak in rural parts of 10 Plains and Western states even though it improved slightly in the past month. Nebraska senators: Death penalty study validates repeal vote By Associated Press • Aug 19, 2016 LINCOLN, Neb. (AP) — Nebraska senators who voted to abolish the death penalty say a recent analysis of the program's costs validates their argument that it's wasteful and ineffective. Do Space offers a Girls Who Code Club By Cheril Lee • Aug 19, 2016 Do Space will offer a Girls Who Code Club starting this fall. Report sets stage for Nebraska session's tax debate By Associated Press • Aug 18, 2016 LINCOLN, Neb. (AP) — A new report says Nebraska could gain a competitive edge over nearby states by reducing its personal and corporate income tax rates while expanding its sales tax and eliminating some business tax credits. Omaha council OKs $250K contract to combat tree-killing bug By Associated Press • Aug 18, 2016 OMAHA, Neb. (AP) — Omaha has committed $250,000 to continue its fight against a tree-killing insect. Former Husker Tagge punished by financial industry watchdog By Associated Press • Aug 18, 2016 OMAHA, Neb. 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Australian firm Champ Private Equity and Hong Kong-based peer Headland Capital Partners have reportedly increased their stakes in vessel company in Miclyn Express Offshore.
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Welcome to the website of the Institute of Geography, University of Bern. Founded in 1886, the Institute comprises Human Geography, Physical Geography, and Integrative Geography and conducts research on all aspects of geography. The Institute offers a Bachelor and a Master program and contributes to several other study programs.
GIUB students at the European Parliament
In the framework of the seminar Political Geography a group of GIUB master's students visited the European Parliament in Strasbourg.


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Then, the error "E_ACT_TOO_MANY_ACTIVATIONS" occurs, indicating that all the authorizations allowed for Adobe Digital Editions on that machine have been used.
Sometimes, you don't receive the error, but run out of activations after switching machines or replacing hardware.
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If possible, generate a System info file before you contact us (see instructions below). Adobe uses this file to help find a common element that's causing this issue for all Adobe Digital Editions users.
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text:
My Mother's Upside Down Cake
Total Time
1hr 25mins
Prep 25 mins
Cook 1 hr
One of the major reasons I joined this website, yes I know there are many recipes. But try this one and see what you think, compare with your own :) You will certainly NOT be disappointed. I also find it is an excuse to get kids to eat fruit! We would always demand this every Sunday after a roast, and pour tonnes of golden syrup over it and let the sponge absorb all the sweet gooeyness.
Ingredients Nutrition
Base of cake
- 8 ounces pineapple (canned and well drained, or fresh)
- 2 ounces butter
- 2 ounces brown sugar
- 1 tablespoon golden syrup or 1 tablespoon honey
- 2 bananas (sliced)
- 5 ounces butter or 5 ounces margarine
- 5 ounces caster sugar
- 3 eggs
- 6 ounces self raising flour
- Preheat the oven to 180-190°C/350-375°F/gas mark 4.
- Prepare the fruit, melt the butter and pour into the bottom of a 7 inch ovenproof dish or cake tin (one with a loose base). Top with the brown sugar and golden syrup and arrange the pineapple rings on the glaze. Add a piece of banana in the centre of each ring (you might need only one banana depending on its size).
- For the Cake: Cream the butter (or margarine) and sugar. Gradually beat in the eggs, then fold in the flour. Spread over the fruit then bake for approximately 1 1/4 hours. Turn out, serve hot with a helping of cool double cream which soaks so yummily into the sponge -- or even cold as a cake rather than a dessert.
- Warning: to not leave the remaining cake batter on the side- you'll demolish it! or the kids will :).
- Serves 4 as a dessert, 6 for tea.
Most Helpful
Very good. Didn't make any changes


text: Day Dreamin' Davey is an adventure game developed by Sculptured Software and released on Nintendo Entertainment System. Continue Reading → Jump to a section: Home Releases Credits Overview Walkthrough Achievements Cheats Day Dreamin' Davey Brainwaves View all → Discussion Article Thought Review 100 Share Day Dreamin' Davey with others Games You May Like Abadox: The Deadly Inner War Adventures of Lolo 3 Alien Hominid Arch Rivals: A Basket Brawl! Day Dreamin' Davey Releases Credits Characters List Overview Walkthrough Achievements Cheats Updates