CorA. Unlike the other transport proteins reported so far, XntAp is dependent on intracellular Ca2+. The transport is also dependent on ΔΨ, but again Mg2+ is not transported to equilibrium, being limited to approximately 0.4 mM free Mg2+ in the cytoplasm. The existence of an intracellular compartment with a much higher free concentration of Mg2+ (8 mM) was supported by the results. Animals The investigation of Mg2+ in animals, including humans, has lagged behind that in bacteria and yeast. This is largely because of the complexity of the systems involved, but also because of the impression within the field that Mg2+ was maintained at high levels in all cells and was unchanged by external influences. Only in the last 25 years has a series of reports begun to challenge this view, with new methodologies finding that free Mg2+ content is maintained at levels where changes might influence cellular metabolism. MRS2 A bioinformatic search of the sequence databases identified one homologue of the MRS2 gene of yeast in a range of metazoans. The protein has a very similar sequence and predicted TM topology to the yeast protein, and the GMN motif is intact at the end of the first TM domain. The human protein, hsaMrs2p, has been localised to the mitochondrial membrane in mouse cells using a GFP fusion protein. Very little is known about the Mg2+ transport characteristics of the protein in mammals, but Zsurka et al. (2001) has shown that the human Mrs2p complements the mrs2 mutants in the yeast mitochondrial Mg2+ uptake system. SLC41 (MgtE) The identification of this gene family in the metazoa began with a signal sequence trap method for isolating secreted and membrane proteins. Much of the identification has come from bioinformatic analyses. Three genes were eventually identified in humans, another three in mouse and three in Caenorhabditis elegans, with a single gene in Anopheles gambiae. The pFAM database lists the MgtE domain as pFAM01769 and additionally identifies a MgtE domain-containing protein in Drosophila melanogaster. The proteins containing the MgtE domain can be divided into seven classes, as defined by pFAM using the type and organisation of the identifiable domains in each protein. Metazoan proteins are present in three of the seven groups. All of the metazoa proteins contain two MgtE domains, but some of these have been predicted only by context recognition (Coin, Bateman and Durbin, unpublished. See the pFAM website for further details).