Metabolomics approach for enzyme discovery

Natsumi Saito, Martin Robert, Sayaka Kitamura, Richard Baran, Tomoyoshi Soga, Hirotada Mori, Takaaki Nishioka, Masaru Tomita

研究成果: Article査読

61 被引用数 (Scopus)

抄録

The search for novel enzymes is an important but difficult task in functional genomics. Here, we present a systematic method based on in vitro assays in combination with metabolite profiling to discover novel enzymatic activities. A complex mixture of metabolites is incubated with purified candidate proteins and the reaction mixture is subsequently profiled by capillary electrophoresis electrospray ionization mass spectrometry (CE-MS). Specific changes in the metabolite composition can directly suggest the presence of an enzymatic activity while subsequent identification of the compounds whose level changed specifically can pinpoint the actual substrate(s) and product(s) of the reaction. We first evaluated the method using several Escherichia coli metabolic enzymes and then applied it to the functional screening of uncharacterized proteins. In this manner, YbhA and YbiV proteins were found to display both phosphotransferase and phosphatase activity toward different sugars/sugar phosphates. Our approach should be broadly applicable and useful for enzyme discovery in any system.

本文言語English
ページ(範囲)1979-1987
ページ数9
ジャーナルJournal of Proteome Research
5
8
DOI
出版ステータスPublished - 2006 8 1

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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