Measurement of enzyme kinetics and inhibitor constants using enthalpy arrays

被引：23

作者：

Rechta, Michael I. ^{[1
]}

Torres, Frank E. ^{[1
]}

De Bruyker, Dirk ^{[1
]}

Bell, Alan G. ^{[1
]}

Klumpp, Martin ^{[2
]}

Bruce, Richard H. ^{[1
]}

机构：

[1] Xerox Corp, Palo Alto Res Ctr, Palo Alto, CA 94304 USA

[2] Novartis Pharma AG, CPC LFP BCS, Novartis Inst Biomed Res Basel, CH-4002 Basel, Switzerland

来源：

ANALYTICAL BIOCHEMISTRY | 2009年 / 388卷 / 02期

基金：

美国国家卫生研究院;

关键词：

Nanocalorimetry; Enzyme assay; Label-free assay; DEPENDENT PROTEIN-KINASE; YEAST HEXOKINASE; SUBSTRATE ACTIVATION; TRYPSIN; CALORIMETRY; PEPTIDES; BINDING; CHYMOTRYPSIN; MECHANISM;

D O I：

10.1016/j.ab.2009.02.028

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Enthalpy arrays enable label-free, solution-based calorimetric detection of molecular interactions in a 96-detector array format. Compared with conventional calorimetry, enthalpy arrays achieve a significant reduction of sample volume and measurement time through the combination of the small size of the detectors and ability to perform measurements in parallel. The current capabilities of the technology for Studying enzyme-catalyzed reactions are demonstrated by determining the kinetic parameters for reactions with three model enzymes. In addition, the technology has been used with two classes of enzymes to determine accurate inhibitor constants for competitive inhibitors from measurements at a single inhibitor concentration. (c) 2009 Elsevier Inc. All rights reserved.

引用

页码：204 / 212

页数：9

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