title: Human cystathionine beta-synthase cDNA: sequence, alternative splicing and expression in cultured cells.
abstract: Cystathionine beta-synthase (CBS) deficiency is the major cause of homocystinuria in humans. The most frequent symptoms of homocystinuria include: dislocated optic lenses, vascular disorders, skeletal abnormalities and mental retardation. Patients with this deficiency have elevated levels of homocyst(e)ine, methionine and low cysteine in their body fluids. These abnormal levels often partially or fully normalize upon treatment with pharmacological doses of vitamin B6. To investigate the molecular and biochemical basis for these conditions, it was necessary to determine the nucleotide and polypeptide sequence of CBS. We report here the human CBS cDNA sequence of 2,554 nucleotides encoding the CBS subunit of 551 amino acids. An intron of 214 bp appears to be retained in the 3'-untranslated region of most of the fibroblast and liver mRNA. We also report a frequent Mspl polymorphism in the 3'-untranslated sequence and two synonymous mutations in the coding region: 699C/T (Y233Y) and 1080C/T (A360A). The amino acid sequence similarity of human and rat CBS is greater than 90%; the enzyme also exhibits 52% similarity to O-acetylserine(thiol)-lyase from bacteria and plants. Lastly, we demonstrate that expression of the human enzyme in CHO cells yields enzymatically active protein of the expected size with a half-life of approximately 14 hrs.
authors.affiliation: Department of Pediatrics, University of Colorado School of Medicine, Denver 80262.
authors.forename: J P
authors.initials: JP
authors.lastname: Kraus
authors.forename: K
authors.initials: K
authors.lastname: Le
authors.forename: M
authors.initials: M
authors.lastname: Swaroop
authors.forename: T
authors.initials: T
authors.lastname: Ohura
authors.forename: T
authors.initials: T
authors.lastname: Tahara
authors.forename: L E
authors.initials: LE
authors.lastname: Rosenberg
authors.forename: M D
authors.initials: MD
authors.lastname: Roper
authors.forename: V
authors.initials: V
authors.lastname: Kozich
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D002478:Cells, Cultured; D006224:Cricetinae; D003541:Cystathionine beta-Synthase / Q000096:biosynthesis / Q000172:deficiency / Q000235:genetics*; D004252:DNA Mutational Analysis; D018076:DNA, Complementary / Q000235:genetics*; D005260:Female; D005347:Fibroblasts / Q000737:chemistry; D006712:Homocystinuria / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000737:chemistry; D008297:Male; D008969:Molecular Sequence Data; D009928:Organ Specificity; D010375:Pedigree; D012150:Polymorphism, Restriction Fragment Length; D012326:RNA Splicing; D051381:Rats / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012867:Skin / Q000737:chemistry; D013045:Species Specificity
chemical_list: D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D003541:Cystathionine beta-Synthase