Diced electrophoresis gel assay for screening enzymes with specified activities

Toru Komatsu, Kenjiro Hanaoka, Alexander Adibekian, Kentaro Yoshioka, Takuya Terai, Tasuku Ueno, Mitsuyasu Kawaguchi, Benjamin F. Cravatt, Tetsuo Nagano

Research output: Contribution to journalArticlepeer-review

21 Citations (Scopus)

Abstract

We have established the diced electrophoresis gel (DEG) assay as a proteome-wide screening tool to identify enzymes with activities of interest using turnover-based fluorescent substrates. The method utilizes the combination of native polyacrylamide gel electrophoresis (PAGE) with a multiwell-plate- based fluorometric assay to find protein spots with the specified activity. By developing fluorescent substrates that mimic the structure of neutrophil chemoattractants, we could identify enzymes involved in metabolic inactivation of the chemoattractants.

Original languageEnglish
Pages (from-to)6002-6005
Number of pages4
JournalJournal of the American Chemical Society
Volume135
Issue number16
DOIs
Publication statusPublished - 2013 Apr 24
Externally publishedYes

ASJC Scopus subject areas

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

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