Abstract
Determining the subcellular location of molecular motors is an essential step in understanding their function and regulation. Although much attention has been focused on the ability of kinesin-like motors to bind reversibly to microtubules, the interaction of this family with cargo has not been extensively studied. Thus, the interactions of kinesin with membrane-bound organelles (MBOs) remains poorly understood, although kinesins are widely thought to serve as motors for movement of MBOs (see also Chapter 12). Methods presented in this chapter were developed to study the regulation of the membrane association of kinesins.
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References
Elluru, R., Bloom, G. S., and Brady, S. T. (1995) Fast axonal transport of kinesin in the rat visual system: functionality of the kinesin heavy chain isoforms. Mol. Biol. Cell 6, 21–40.
Stenoien, D. S. and Brady, S. T. (1997) Immunochemical analysis of kinesin light chain function. Mol. Biol. Cell 8, 675–689.
Pfister, K. K., Wagner, M. C., Stenoien, D., Bloom, G. S., and Brady, S. T. (1989) Monoclonal antibodies to kinesin heavy and light chains stain vesicle-like structures, but not microtubules, in cultured cells. J. Cell Biol. 108, 1453–1463.
Wagner, M. C., Pfister, K. K., Bloom, G. S., and Brady, S. T. (1989) Copurification of kinesin polypeptides with microtubule-stimulated Mg-ATPase activity and kinetic analysis of enzymatic processes. Cell Motil. Cytoskeleton 12, 195–215.
Tsai, M.-Y., Morfini, G., Szebenyi, G., and Brady, S. T. (1999) Release of kinesin from vesicles by hsc70 and regulation of fast axonal transport. Mol. Biol. Cell 11, 2161–2173.
Goslin, K. and Banker, G. (1991) Rat hippocampal neurons in low-density culture, in Culturing Nerve Cells (Banker, G. and K. Goslin, eds.), MIT Press, Cambridge, MA, pp. 252–281.
Ramsby, M. L. and Makowski, G. S. (1998) Differential detergent fractionation of eukaryotic cells. Analysis by two dimensional gel electrophoresis, in 2-D Proteome Analysis Protocols (Link, A. J., ed.), Humana, Totowa, NJ, pp. 53–65.
Ramsby, M. L., Makowski, G. S., and Khairallah, E. A. (1994) Differential detergent fractionation of isolated hepatocytes: biochemical, immunochemical, and two dimensional gel electrophoresis characterization of cytoskeletal compartments. Electrophoresis 15, 265–277.
Carter, N. (1997) Permeabilization strategies to study protein phosphorylation, in Current Protocols in Protein Science (Chanda, V. B., ed.), Wiley, New York.
Elluru, R., Bloom, G. S., and Brady, S. T. (1995) Fast axonal transport of kinesin in the rat visual system: functionality of the kinesin heavy chain isoforms. Mol. Biol. Cell 6, 21–40.
Rodriguez, L., Stirling, C. J., and Woodman, P. G. (1994) Multiple N-ethylmaleimidesensitive components are required for endosomal vesicle fusion. Mol. Biol. Cell 5, 773–783.
Pfister, K. K., Wagner, M. C., Stenoien, D., Bloom, G. S., and Brady, S. T., (1989) Monoclonal antibodies to kinesin heavy and light chains stain vesicle-like structures, but not microtubules, in cultured cells. J. Cell Biol. 108, 1453–1463.
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Morfini, G., Tsai, MY., Szebenyi, G., Brady, S.T. (2001). Approaches to Study Interactions Between Kinesin Motors and Membranes. In: Vernos, I. (eds) Kinesin Protocols. Methods in Molecular Biology™, vol 164. Humana Press. https://doi.org/10.1385/1-59259-069-1:147
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DOI: https://doi.org/10.1385/1-59259-069-1:147
Publisher Name: Humana Press
Print ISBN: 978-0-89603-766-3
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