TY - JOUR
T1 - Diced electrophoresis gel assay for screening enzymes with specified activities
AU - Komatsu, Toru
AU - Hanaoka, Kenjiro
AU - Adibekian, Alexander
AU - Yoshioka, Kentaro
AU - Terai, Takuya
AU - Ueno, Tasuku
AU - Kawaguchi, Mitsuyasu
AU - Cravatt, Benjamin F.
AU - Nagano, Tetsuo
PY - 2013/4/24
Y1 - 2013/4/24
N2 - 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.
AB - 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.
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U2 - 10.1021/ja401792d
DO - 10.1021/ja401792d
M3 - Article
C2 - 23581642
AN - SCOPUS:84876721515
SN - 0002-7863
VL - 135
SP - 6002
EP - 6005
JO - Journal of the American Chemical Society
JF - Journal of the American Chemical Society
IS - 16
ER -