title: Cloning, expression and characterization of human thioltransferase (glutaredoxin) in E. coli.
abstract: PCR primers were designed from the known amino acid (aa) sequence for human red blood cell thioltransferase (hRBC TTase) and the known cDNA sequence for pig liver TTase (82% homologous) and used to amplify thioltransferase from a pool of human brain cDNAs. The PCR product was inserted into the pKK233-2 expression vector. The DNA sequence of the insert agreed with the aa sequence. High level expression of the enzyme was accomplished in E. coli, and Western blot analysis confirmed its identity. Recombinant TTase displayed catalytic properties indistinguishable from natural hRBC TTase.
authors.affiliation: Department of Pharmacology, Case Western Reserve University School of Medicine, Cleveland, OH 44106, USA.
authors.forename: C A
authors.initials: CA
authors.lastname: Chrestensen
authors.forename: C B
authors.initials: CB
authors.lastname: Eckman
authors.forename: D W
authors.initials: DW
authors.lastname: Starke
authors.forename: J J
authors.initials: JJ
authors.lastname: Mieyal
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002384:Catalysis; D003001:Cloning, Molecular; D004912:Erythrocytes / Q000201:enzymology; D004926:Escherichia coli; D015870:Gene Expression; D054477:Glutaredoxins; D006801:Humans; D008969:Molecular Sequence Data; D010088:Oxidoreductases / Q000235:genetics* / Q000378:metabolism; D011490:Protein Disulfide Reductase (Glutathione); D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D013552:Swine
chemical_list: D054477:Glutaredoxins; D011993:Recombinant Fusion Proteins; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione)