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J Am Chem Soc. 2013 Apr 24;135(16):6002-5. doi: 10.1021/ja401792d. Epub 2013 Apr 16.

Diced electrophoresis gel assay for screening enzymes with specified activities.

Author information

1
Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.

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.

PMID:
23581642
DOI:
10.1021/ja401792d
[Indexed for MEDLINE]

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