A non-radioactive assay for selenophosphate synthetase activity using recombinant pyruvate pyrophosphate dikinase from Thermus thermophilus HB8

作者：Kamada Saho; Okugochi Takahiro; Asano Kaori; Tobe Ryuta; Mihara Hisaaki; Nemoto Michiko; Inagaki Kenji; Tamura Takashi^*

来源：Bioscience Biotechnology and Biochemistry, 2016, 80(10): 1970-1972.

DOI：10.1080/09168451.2016.1200458

• selenophosphate synthetase
• monoselenophosphate
• pyruvate pyrophosphate dikinase
• Thermus thermophilus HB8
• non-RI method

Biosynthesis of selenocysteine-containing proteins requires monoselenophosphate, a selenium-donor intermediate generated by selenophosphate synthetase (Sephs). A non-radioactive assay was developed as an alternative to the standard [8-C-14] AMP-quantifying assay. The product, AMP, was measured using a recombinant pyruvate pyrophosphate dikinase from Thermus thermophilus HB8. The K-M and k(cat) for Sephs2-Sec60Cys were determined to be 26M and 0.352min(-1), respectively.

• 出版日期2016