title: A novel 9-bp insertion detected in steroid 21-hydroxylase gene (CYP21A2): prediction of its structural and functional implications by computational methods.
abstract: BACKGROUND
  Steroid 21-hydroxylase deficiency is the most common cause of congenital adrenal hyperplasia (CAH). Detection of underlying mutations in CYP21A2 gene encoding steroid 21-hydroxylase enzyme is helpful both for confirmation of diagnosis and management of CAH patients. Here we report a novel 9-bp insertion in CYP21A2 gene and its structural and functional consequences on P450c21 protein by molecular modeling and molecular dynamics simulations methods.
  
  
  METHODS
  A 30-day-old child was referred to our laboratory for molecular diagnosis of CAH. Sequencing of the entire CYP21A2 gene revealed a novel insertion (duplication) of 9-bp in exon 2 of one allele and a well-known mutation I172N in exon 4 of other allele. Molecular modeling and simulation studies were carried out to understand the plausible structural and functional implications caused by the novel mutation.
  
  
  RESULTS
  Insertion of the nine bases in exon 2 resulted in addition of three valine residues at codon 71 of the P450c21 protein. Molecular dynamics simulations revealed that the mutant exhibits a faster unfolding kinetics and an overall destabilization of the structure due to the triple valine insertion was also observed.
  
  
  CONCLUSIONS
  The novel 9-bp insertion in exon 2 of CYP21A2 genesignificantly lowers the structural stability of P450c21 thereby leading to the probable loss of its function.
authors.affiliation: Department of Molecular Endocrinology, National Institute for Research in Reproductive Health, Indian Council of Medical Research, J M Street, Parel, Mumbai, Maharashtra, India. dsudhisha@yahoo.com
authors.forename: Sudhisha
authors.initials: S
authors.lastname: Dubey
authors.forename: Susan
authors.initials: S
authors.lastname: Idicula-Thomas
authors.forename: Mohammad
authors.initials: M
authors.lastname: Anwaruddin
authors.forename: Chinnaraj
authors.initials: C
authors.lastname: Saravanan
authors.forename: R Raveendra
authors.initials: RR
authors.lastname: Varma
authors.forename: Anurupa
authors.initials: A
authors.lastname: Maitra
mesh_terms: D000312:Adrenal Hyperplasia, Congenital / Q000201:enzymology / Q000235:genetics; D000483:Alleles; D000595:Amino Acid Sequence; D001483:Base Sequence; D019295:Computational Biology / Q000379:methods*; D004252:DNA Mutational Analysis; D005091:Exons; D005260:Female; D006801:Humans; D007223:Infant; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D011487:Protein Conformation; D013255:Steroid 21-Hydroxylase / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
chemical_list: D013255:Steroid 21-Hydroxylase