Shikata H, Koyama S, Egi Y, Yamada K, Kawasaki T
Department of Biochemistry, Hiroshima University School of Medicine, Japan.
Biochem Int. 1989 May;18(5):933-41.
An attempt was made to purify a porcine skeletal muscle enzyme catalyzing the formation of thiamin triphosphate (TTP) from thiamin diphosphate (TDP), requiring ATP, Mg2+ and a cofactor (creatine). As the purification proceeded, the reaction requirements for ATP and creatine were lost and then a requirement for ADP was manifested. The activity responsible for TTP synthesis from TDP, ADP, and Mg2+ was found to be copurified with adenylate kinase [EC 2.7.4.3] activity, and was finally purified to a single band on SDS-PAGE. Antiserum obtained against the purified enzyme preparation inhibited both adenylate kinase activity and the TTP-synthesizing activity to exactly the same extent. These results indicate that adenylate kinase catalyzes TTP formation from TDP in vitro.
人们试图纯化一种猪骨骼肌酶，该酶可催化由硫胺二磷酸（TDP）形成硫胺三磷酸（TTP），反应需要ATP、Mg2+和一种辅因子（肌酸）。随着纯化过程的进行，对ATP和肌酸的反应需求消失，随后表现出对ADP的需求。发现负责从TDP、ADP和Mg2+合成TTP的活性与腺苷酸激酶[EC 2.7.4.3]活性共纯化，最终在SDS-PAGE上纯化到单一条带。针对纯化的酶制剂获得的抗血清对腺苷酸激酶活性和TTP合成活性的抑制程度完全相同。这些结果表明，腺苷酸激酶在体外催化从TDP形成TTP。
