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J. Biol. Chem., Vol. 279, Issue 38, 40100-40111, September 17, 2004
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From the
Department of Neurology, University of Alabama, Birmingham, Alabama 35294 and the ¶Department of Physiology, University of Pennsylvania, Philadelphia, Pennsylvania 19104
Cytoplasmic transport is mediated by a group of molecular motors that typically work in isolation, under conditions where they must move their cargos long distances without dissociating from their tracks. This processive behavior requires specific adaptations of motor enzymology to meet these unique physiologic demands. One of these involves the ability of the two heads of a processive motor to communicate their structural states to each other. In this study, we examine a processive motor from the myosin superfamily myosin V. We have measured the kinetics of nucleotide release, of phosphate release, and of the weak-to-strong transition, as this motor interacts with actin, and we have used these studies to develop a model of how myosin V functions as a transport motor. Surprisingly, both heads release phosphate rapidly upon the initial encounter with an actin filament, suggesting that there is little or no intramolecular strain associated with this step. However, ADP release can be affected by both forward and rearward strain, and under steady-state conditions it is essentially prevented in the lead head until the rear head detaches. Many of these features are remarkably like those underlying the processive movement of kinesin on microtubules, supporting our hypothesis that different molecular motors satisfy the requirement for processive movement in similar ways, regardless of their particular family of origin.
Received for publication, March 8, 2004 , and in revised form, June 28, 2004.
* This work was supported by National Institutes of Health Grants AR048565 (to S. S. R.) and AR35661 (to H. L. S.). The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Neurology, University of Alabama at Birmingham, FOT 1020, 1530 3rd Ave. S., Birmingham, AL 35294. Tel.: 205-934-0284; Fax: 205-975-7546; E-mail: stevensr{at}uab.edu.
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