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The ABRF-MIRG'02 Study: Assembly State, Thermodynamic, and Kinetic Analysis of an Enzyme/Inhibtor Interaction

Published

Author(s)

D Myszka, Huixin He, F Arisaka, O Byron, Edward Eisenstein, P Hensley, J Thomson, C Lombardo, Frederick P. Schwarz, W Stafford, M Doyle

Abstract

Fully characterizing the interactions involving biomolecules requires information on the assembly state, affinity, kinetics, and thermodynamics associated with complex formation. The analytical technologies often utilized to measure biomolecular interactions include analytical ultracentrifugation (AUC), isothermal titration calorimetry (ITC), and surface plasmon resonance (SPR). In order to evaluate the capabilities of Core Facilities to implement these technologies, the Molecular Interactions Research Group (MIREG) of the Association of Biomolecular Resource Facilities (ABRF) developed a standardized model system and distributed it to a panel of AUC, ITC, and SPR operators. The model system was composed of a well-characterized enzyme/inhibitor pair, namely bovine carbonic anhydrase II (CA II) and 4-carboxybenzenesulfonamide (CBS). Study participants were asked to measure one or more of the following: 1) the molecular mass, homogeneity, and assembly state of CA II by AUC; 2) the affinity and thermodynamics for complex formation by ITC; and 3) the affinity and kinetics of complex formation by SPR. The results from this study provide a benchmark for comparing the capabilities of individual laboratories and defining the utility of the different instrumentation.
Citation
Journal of Biomolecular Techniques
Volume
14 No. 4

Keywords

analytical, calorimetry, carbonic anhydrase, isothermal titration, surface plasmon resonance, ultracentrifugation

Citation

Myszka, D. , He, H. , Arisaka, F. , Byron, O. , Eisenstein, E. , Hensley, P. , Thomson, J. , Lombardo, C. , Schwarz, F. , Stafford, W. and Doyle, M. (2003), The ABRF-MIRG'02 Study: Assembly State, Thermodynamic, and Kinetic Analysis of an Enzyme/Inhibtor Interaction, Journal of Biomolecular Techniques (Accessed April 14, 2024)
Created November 30, 2003, Updated October 12, 2021