title: In silico analysis of five missense mutations in CYP1B1 gene in Pakistani families affected with primary congenital glaucoma.
abstract: OBJECTIVE
  The purpose of this study was to characterize the five missense mutations in CYP1B1 gene identified in Pakistani families affected with primary congenital glaucoma (PCG) using various bioinformatics and protein modeling tools.
  
  
  METHODS
  We previously reported four novel missense mutations in CYP1B1 gene segregating in consanguineous Pakistani families. These mutations were identified by direct sequencing of all coding exons, the exon-intron boundaries and the 5' untranslated region of CYP1B1 using genomic DNA from affected and unaffected family members. In order to understand the effect of CYP1B1 mutations on protein structure and function, we used bioinformatics tools to investigate five mutations including four novels (W434R, D374E, L487P and L177R) and one known (E229K) mutation previously reported by our group in four Pakistani PCG-affected families.
  
  
  RESULTS
  In silico analysis of the missense mutations using the computational algorithms SNAP, I-Mutant 2.0 IUPred, PrDOS and PASTA predicted pathogenic effects on stability and function of protein.
  
  
  CONCLUSIONS
  In silico analysis of identified mutations confirmed their molecular pathogenicity. Similar analysis will be helpful in understanding of the biological role of CYP1B1 and the effect of mutations on the regulatory and enzymatic functions of CYP1B1 that result in PCG.
authors.affiliation: Department of Animal Sciences, Faculty of Biological Sciences, Quaid-i-Azam University, Islamabad, Pakistan.
authors.forename: Sabika
authors.initials: S
authors.lastname: Firasat
authors.affiliation: Genetics Division, Department of Livestock Production, University of Veterinary and Animal Sciences, Ravi Campus, Pattoki, Pakistan. haibakaul@gmail.com.
authors.forename: Haiba
authors.initials: H
authors.lastname: Kaul
authors.affiliation: Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan.
authors.forename: Usman Ali
authors.initials: UA
authors.lastname: Ashfaq
authors.affiliation: Department of Bioinformatics and Biotechnology, Government College University, Faisalabad, Pakistan.|School of Biotechnology and Biomolecular Sciences, The University of New South Wales, Kensington, Sydney, Australia.
authors.forename: Sobia
authors.initials: S
authors.lastname: Idrees
mesh_terms: D065633:Cytochrome P-450 CYP1B1 / Q000235:genetics*; D004252:DNA Mutational Analysis; D005260:Female; D005901:Glaucoma / Q000151:congenital / Q000235:genetics*; D006801:Humans; D008297:Male; D020125:Mutation, Missense; D010154:Pakistan; D010375:Pedigree
chemical_list: C585496:CYP1B1 protein, human; D065633:Cytochrome P-450 CYP1B1
keywords: Bioinformatics; CYP1B1; Missense mutation; Primary congenital glaucoma (PCG)