title: Properties of the thiamin triphosphate-synthesizing activity catalyzed by adenylate kinase (isoenzyme 1).
abstract: Adenylate kinase isozyme 1 (AK1) catalyzes thiamin triphosphate (TTP) formation from thiamin diphosphate (TDP) and ADP. The properties of the TTP-synthesizing activity of purified AK1 from porcine skeletal muscle were studied. The activity was found to require TDP, ADP, and Mg2+, and ATP was only 14.4% as active as ADP. Thiamin monophosphate (TMP) and thiamin were not utilized as substrates. ADP was specific as a phosphate donor; and CDP, UDP, and GDP supported TTP formation at rates less than 1% of that with ADP. Optimal pH and temperature for the TTP-synthesizing activity were 10.0 and 37 degrees C, respectively. The activity showed saturation kinetics for both substrates, and the Km values for TDP and ADP were calculated to be 0.83 mM and 43 microM, respectively. The enzyme catalyzed the reverse reaction (TTP + AMP----TDP + ADP) and stoichiometry between TTP and TDP was demonstrated in the forward and reverse reactions.
authors.affiliation: Department of Biochemistry, Hiroshima University School of Medicine, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Shikata
authors.forename: Y
authors.initials: Y
authors.lastname: Egi
authors.forename: S
authors.initials: S
authors.lastname: Koyama
authors.forename: K
authors.initials: K
authors.lastname: Yamada
authors.forename: T
authors.initials: T
authors.lastname: Kawasaki
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000263:Adenylate Kinase / Q000378:metabolism*; D000818:Animals; D002384:Catalysis; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000378:metabolism; D007700:Kinetics; D010770:Phosphotransferases / Q000378:metabolism*; D013552:Swine; D013696:Temperature; D013831:Thiamine / Q000031:analogs & derivatives*; D013838:Thiamine Triphosphate / Q000096:biosynthesis*
chemical_list: D007527:Isoenzymes; D013838:Thiamine Triphosphate; D000244:Adenosine Diphosphate; D010770:Phosphotransferases; D000263:Adenylate Kinase; D013831:Thiamine