Single-enzyme kinetics of CALB-catalyzed hydrolysis

被引:155
作者
Velonia, K [1 ]
Flomenbom, O
Loos, D
Masuo, S
Cotlet, M
Engelborghs, Y
Hofkens, J
Rowan, AE
Klafter, J
Nolte, RJM
de Schryver, FC
机构
[1] Raboud Univ Nijmegen, Dept Organ Chem, Inst Mol & Mat, NL-6525 ED Nijmegen, Netherlands
[2] Katholieke Univ Leuven, Dept Chem, B-3001 Heverlee, Belgium
[3] Tel Aviv Univ, Sackler Fac Exact Sci, Sch Chem, IL-69978 Tel Aviv, Israel
[4] Dept Biochem, Lab Chem & Biol Dynam, B-3001 Heverlee, Belgium
关键词
analytical methods; enzymes; fluorescence; microscopy; kinetics; single-molecule studies;
D O I
10.1002/anie.200460625
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
Real-time measurement of the catalysis and substrate kinetics of a single-enzyme hydrolysis reaction is demonstrated with confocal fluorescence microscopy (CFM; see picture, green = CFM beam). A single lipase is shown to have a broad range of conformations; each conformation contributes to the overall enzymatic activity, an observation that is often masked by ensemble measurements.
引用
收藏
页码:560 / 564
页数:5
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