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Kinetic and Equilibrium Analysis of the Myosin ATPase

Overview
Journal Methods Enzymol
Specialty Biochemistry
Date 2009 Mar 18
PMID 19289206
Citations 96
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Abstract

The myosin superfamily consists of more than 35 classes (each consisting of multiple isoforms) that have diverse cellular activities. The reaction pathway of the actin-activated myosin ATPase appears to be conserved for all myosin isoforms, but the rate and equilibrium constants that define the ATPase pathway vary significantly across the myosin superfamily, resulting in kinetic differences that that allow myosins to carry out diverse mechanical functions. Therefore, it is important to determine the lifetimes and relative populations of the key biochemical intermediates to obtain an understanding of a particular myosin's cellular function. This chapter provides procedures for determining the overall and individual rate and equilibrium constants of the actomyosin ATPase cycle, including actomyosin binding and dissociation, ATP binding, ATP hydrolysis, phosphate release, and ADP release and binding. Many of the methods described in the chapter are applicable to the characterization of other ATPase enzymes.

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