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

Saho Kamada, Takahiro Okugochi, Kaori Asano, Ryuta Tobe, Hisaaki Mihara, Michiko Nemoto, Kenji Inagaki, Takashi Tamura

Research output: Contribution to journalArticle

Abstract

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 [8-14 C] AMP-quantifying assay. The product, AMP, was measured using a recombinant pyruvate pyrophosphate dikinase from Thermus thermophilus HB8. The KM and kcat for Sephs2-Sec60Cys were determined to be 26 μM and 0.352 min-1 , respectively.

Original languageEnglish
Pages (from-to)1970-1972
Number of pages3
JournalBioscience, Biotechnology and Biochemistry
Volume80
Issue number10
DOIs
Publication statusPublished - 2016

Keywords

  • Monoselenophosphate
  • Non-RI method
  • Pyruvate pyrophosphate dikinase
  • Selenophosphate synthetase
  • Thermus thermophilus hb8

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biotechnology
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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