title: Three dimensional structure of human biotinidase: computer modeling and functional correlations.
abstract: Untreated individuals with deficient activity of biotinidase, the enzyme responsible for recycling the vitamin biotin, usually exhibit neurological and cutaneous findings. To better understand the variability in expression of the disorder it is important to understand the structure of the enzyme and the putative effects of various mutations on its activity. Past attempts to express and purify sufficient quantities of the enzyme by us and others have failed. Therefore, we have resorted to computer modeling using homologous related, crystallized nitrilases/amidases to predict the 3-dimensional structure of biotinidase. The resultant structure is a two domain protein with the catalytic triad consisting of glutamate, lysine and cysteine, within the larger domain. The model predicts multiple glycosylation sites at the surface of the enzyme and multiple disulfide bonds. The precise location of the biotin-binding site could not be determined. Characteristics of 45 missense mutations known to cause profound and partial biotinidase deficiency were examined, including their location, their distance from the catalytic triad, and their potential effect on the structure of the enzyme. Although there are obviously short-comings in predicting the 3-dimensional structure of a protein without crystallographic data, because of the marked homology between biotinidase and specific crystallized amidases/nitrilases, the predicted 3-dimensional structure of biotinidase is probable and should be useful providing clues to structure-function relationships and ultimately the effect of mutations on altering the enzyme's hydrolase and transferase activities.
authors.affiliation: Department of Medical Genetics, Henry Ford Hospital, and Center for Molecular Medicine and Genetics, Wayne State University, Detroit, MI 48202, USA.
authors.forename: Kirit
authors.initials: K
authors.lastname: Pindolia
authors.forename: Kevin
authors.initials: K
authors.lastname: Jensen
authors.forename: Barry
authors.initials: B
authors.lastname: Wolf
mesh_terms: D000595:Amino Acid Sequence; D043523:Biotinidase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003198:Computer Simulation; D017124:Conserved Sequence; D016208:Databases, Factual; D006801:Humans; D008969:Molecular Sequence Data; D020125:Mutation, Missense; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid
chemical_list: D043523:Biotinidase