Abstract
Methods for assay of ATPase rates can be classified into broad categories depending on whether an ATP-regenerating system or a continuous coupled assay is employed versus quenching at discrete times for subsequent analysis for production of Pi or ADP (or, more rarely, for the loss of ATP).
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References
Sumner, J. B. (1944) A method for the colorimetric analysis of phosphorus. Science 100, 413–414.
Hackney, D. D., Stempel, K. E., and Boyer, P. D. (1980) Oxygen-18 probes of enzymic reactions of phosphate compounds. Methods Enzymol. 64, 60–83.
Hackney, D. D. and Clark, P. K. (1985) Steady state kinetics at high enzyme concentration: the myosin ATPase. J. Biol. Chem. 260, 5505–5510.
Penney, C. L. and Bolger, G. (1978) A simple microassay for inorganic phosphate, II. Anal. Biochem. 89, 297–303.
Jiang, W. and Hackney, D. D. (1997) Monomeric kinesin head domains hydrolyze multiple ATP molecules before release from a microtubule. J. Biol. Chem. 272, 5616–5621.
Williams, R. C., Jr. and Lee, J. C. (1982) Preparation of tubulin from brain. Methods Enzymol. 85, 376–385.
Huang, T.-G. and Hackney, D. D. (1994) Drosophila kinesin minimal motor domain expressed in Escherichia coli. Purification and kinetic characterization. J. Biol. Chem. 269, 16,493–16,501.
Hackney, D. D. (1994) The rate limiting step in microtubule-stimulated ATP hydrolysis by dimeric kinesin head domains occurs while bound to the microtubule. J. Biol. Chem. 269, 16,508–16,511.
Hackney, D. D., Levitt, J. D., and Suhan, J. (1991) Kinesin undergoes a 9 S to 6 S conformational transition. J. Cell Biol. 115, 388a.
Stock, M. F., Guerrero, J., Cobb, B., Eggers, C. T., Huang, T-G., Li, X., and Hackney, D. D. (1999) Formation of the compact conformer of kinesin requires a C-terminal heavy chain domain and inhibits microtubule-stimulated ATPase activity. J. Biol. Chem. 274, 14,617–14,623.
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© 2001 Humana Press Inc.
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Hackney, D.D., Jiang, W. (2001). Assays for Kinesin Microtubule-Stimulated AT Pase Activity. In: Vernos, I. (eds) Kinesin Protocols. Methods in Molecular Biology™, vol 164. Humana Press. https://doi.org/10.1385/1-59259-069-1:65
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DOI: https://doi.org/10.1385/1-59259-069-1:65
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