Kinetic controlled affinity labeling of target enzyme with thioester chemistry.
High specificity has been an important feature in affinity labeling for target profiling. Especially, to label targets via rapidly progressing reactions with consumption of ligand (probe), high specificity of reaction with common functional groups of target protein should be achieved without reactions with similar groups of non-target proteins. Herein, we demonstrate the kinetic controlled affinity labeling of acyl CoA synthetase using a fatty acid analogue containing a phenylthioester linkage. High specificity was attained by accelerating the labeling rate in the binding pocket. This approach could be useful for profiling a series of target enzymes and transporters in signal transduction pathways.
Laboratory of Biorecognition Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2603 Sugitani, Toyama 930-0194, Japan. Electronic address: ttomo@pha.u-toyama.ac.jp.
Takenori
T
Tomohiro
Laboratory of Biorecognition Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2603 Sugitani, Toyama 930-0194, Japan.
Masahiro
M
Nakabayashi
Laboratory of Biorecognition Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2603 Sugitani, Toyama 930-0194, Japan.
Yuka
Y
Sugita
Laboratory of Biorecognition Chemistry, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2603 Sugitani, Toyama 930-0194, Japan.
Shota
S
Morimoto
D000345:Affinity Labels / Q000737:chemistry*; D001710:Biotin / Q000737:chemistry; D003066:Coenzyme A Ligases / Q000737:chemistry*; D004952:Esters / Q000737:chemistry; D007700:Kinetics; D019814:Myristic Acid / Q000737:chemistry; D019308:Palmitic Acid / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry*
D000345:Affinity Labels; D004952:Esters; D012709:Serum Albumin; D013438:Sulfhydryl Compounds; D019814:Myristic Acid; D019308:Palmitic Acid; D001710:Biotin; D003066:Coenzyme A Ligases
Activity-based affinity labeling; Acyl CoA synthetase; Kinetic; Thioester