Isothermal titration calorimetry: experimental design, data analysis, and probing macromolecule/ligand binding and kinetic interactions. Export

@article{Freyer2008, abstract = {Isothermal titration calorimetry (ITC) is now routinely used to directly characterize the thermodynamics of biopolymer binding interactions and the kinetics of enzyme-catalyzed reactions. This is the result of improvements in ITC instrumentation and data analysis software. Modern ITC instruments make it possible to measure heat effects as small as 0.1 microcal (0.4 microJ), allowing the determination of binding constants, K's, as large as 10(8) - 10(9)M(-1). Modern ITC instruments make it possible to measure heat rates as small as 0.1 microcal/sec, allowing for the precise determination of reaction rates in the range of 10(-12) mol/sec. Values for K(m) and k(cat), in the ranges of 10(-2) - 10(3) microM and 0.05 - 500 sec(-1), respectively, can be determined by ITC. This chapter reviews the planning of an optimal ITC experiment for either a binding or kinetic study, guides the reader through simulated sample experiments, and reviews analysis of the data and the interpretation of the results.}, author = {Freyer, M. W. and Lewis, E. A.}, citeulike-article-id = {3888993}, citeulike-linkout-0 = {http://dx.doi.org/10.1016/S0091-679X(07)84004-0}, citeulike-linkout-1 = {http://view.ncbi.nlm.nih.gov/pubmed/17964929}, citeulike-linkout-2 = {http://www.hubmed.org/display.cgi?uids=17964929}, doi = {10.1016/S0091-679X(07)84004-0}, issn = {0091-679X}, journal = {Methods in cell biology}, keywords = {itc, protein-ligand-binding}, pages = {79--113}, posted-at = {2009-01-15 15:01:23}, priority = {4}, title = {Isothermal titration calorimetry: experimental design, data analysis, and probing macromolecule/ligand binding and kinetic interactions.}, url = {http://dx.doi.org/10.1016/S0091-679X(07)84004-0}, volume = {84}, year = {2008} }

TY - JOUR ID - Freyer2008 L3 - citeulike-article-id:3888993 N2 - Isothermal titration calorimetry (ITC) is now routinely used to directly characterize the thermodynamics of biopolymer binding interactions and the kinetics of enzyme-catalyzed reactions. This is the result of improvements in ITC instrumentation and data analysis software. Modern ITC instruments make it possible to measure heat effects as small as 0.1 microcal (0.4 microJ), allowing the determination of binding constants, K's, as large as 10(8) - 10(9)M(-1). Modern ITC instruments make it possible to measure heat rates as small as 0.1 microcal/sec, allowing for the precise determination of reaction rates in the range of 10(-12) mol/sec. Values for K(m) and k(cat), in the ranges of 10(-2) - 10(3) microM and 0.05 - 500 sec(-1), respectively, can be determined by ITC. This chapter reviews the planning of an optimal ITC experiment for either a binding or kinetic study, guides the reader through simulated sample experiments, and reviews analysis of the data and the interpretation of the results. JF - Methods in cell biology SN - 0091-679X EP - 113 TI - Isothermal titration calorimetry: experimental design, data analysis, and probing macromolecule/ligand binding and kinetic interactions. VL - 84 SP - 79 KW - itc KW - protein-ligand-binding AU - Freyer, MW AU - Lewis, EA PY - 2008/// UR - http://dx.doi.org/10.1016/S0091-679X(07)84004-0 ER -