title: Human serum biotinidase. cDNA cloning, sequence, and characterization.
abstract: Biotinidase (EC 3.5.1.12) catalyzes the hydrolysis of biocytin, the product of biotin-dependent carboxylase degradation, to biotin and lysine. Biotinidase deficiency is an inherited metabolic disorder of biotin recycling that is characterized by neurological and cutaneous abnormalities, and can be successfully treated with biotin supplementation. Sequences of tryptic peptides of the purified human serum enzyme were used to design oligonucleotide primers for polymerase chain reaction amplification from human hepatic total RNA to generate putative biotinidase cDNA fragments. Sequence analysis of a cDNA isolated from a human liver library by plaque hybridization with the largest cDNA probe revealed an open reading frame of 1629 bases encoding a protein of 543 amino acid residues, including 41 amino acids of a potential signal peptide. Comparison of the open reading frame with the known biotinidase tryptic peptides and recognition of the expressed protein encoded by this cDNA by monoclonal antibodies prepared against purified biotinidase demonstrated the identity of this cDNA. Southern analyses suggested that biotinidase is a single copy gene and revealed that human cDNA probes hybridized to genomic DNA from mammals, but not from chicken or yeast. Northern analysis indicated the presence of biotinidase mRNA in human heart, brain, placenta, liver, lung, skeletal muscle, kidney, and pancreas.
authors.affiliation: Department of Human Genetics, Medical College of Virginia/Virginia Commonwealth University, Richmond 23298.
authors.forename: H
authors.initials: H
authors.lastname: Cole
authors.forename: T R
authors.initials: TR
authors.lastname: Reynolds
authors.forename: J M
authors.initials: JM
authors.lastname: Lockyer
authors.forename: G A
authors.initials: GA
authors.lastname: Buck
authors.forename: T
authors.initials: T
authors.lastname: Denson
authors.forename: J E
authors.initials: JE
authors.lastname: Spence
authors.forename: J
authors.initials: J
authors.lastname: Hymes
authors.forename: B
authors.initials: B
authors.lastname: Wolf
mesh_terms: D000581:Amidohydrolases / Q000096:biosynthesis / Q000097:blood* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D043523:Biotinidase; D015152:Blotting, Northern; D003001:Cloning, Molecular; D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D017931:DNA Primers; D015342:DNA Probes; D018076:DNA, Complementary; D010446:Peptide Fragments; D012313:RNA; D000581:Amidohydrolases; D043523:Biotinidase