PMC7265266_01
Pedigree and identification of the mutation in the family. (a) The segregation and genotyping analyses of the family members concerning the (NM_000104 (CYP1B1):c.1099_1099delG) mutation. (b) The Sanger sequencing method confirmed the mutation c.1099_1099delG in the patient and her parents. The deleted nucleotide was designated in the bold and red font within the reference sequence (wild-type) and also pointed by an arrow in the electropherogram. The Sanger sequencing was performed by the reverse primer; then the base-calling errors appeared next to the mutation position and represent different reads in heterozygous parents
JCO-32-114-g001.jpg
PMC7265266_01_JCO-32-114-g001.jpg
F1
The subject was born of consanguineous marriage [Figure 1a]. The Sanger sequencing and segregation analysis confirmed homozygosity of the variant in the patient, while her parents were heterozygote [Figure 1a and b].
The three-dimensional structure prediction and pairwise alignment of the protein in the mutant and wild-type forms. (a) The three-dimensional structure prediction of the wild-type form of the cytochrome P450 family 1 subfamily B member 1 (CYP1B1) protein (right view), while aspartic acid, at the position 367, is located at the protein surface and colored green. (b) The three-dimensional structure prediction of the mutant form of the CYP1B1 protein (right view) in which threonine, at the position 367, is located at the protein surface and colored green. (c) The wild-type protein was colored as green, while the mutant one was colored red. The dashed red circle encompassed the un-aligned features of the two forms of the CYP1B1 protein (wild-type vs. mutant) and represented the structural changes caused by p.(D367Tfs*61) mutation
JCO-32-114-g002.jpg
PMC7265266_01_JCO-32-114-g002.jpg
F2
The results of the 3D structure prediction for both mutant and wild-type forms of the CYP1B1 protein revealed that some features might be affected in the mutant form of the protein [Figure 2a and b]. It was also demonstrated that several protein features of the mutant form of CYP1B1 would be lost in each output pairwise alignment when compared with its reference alignment [Figure 2c].