期刊名称：Proceedings of the National Academy of Sciences
印刷版ISSN：0027-8424
电子版ISSN：1091-6490
出版年度：2014
卷号：111
期号：38
页码：E3986-E3995
DOI：10.1073/pnas.1406535111
语种：English
出版社：The National Academy of Sciences of the United States of America
摘要：SignificanceIntracellular cargo transport is carried out by ensembles of cytoskeleton-based molecular motors, such as myosin Va. Physiologically, motor molecules are bound to (and mechanically coupled through) the vesicular membrane, which is a fluid lipid bilayer. Utilizing a combination of experiment and computer simulation, we characterize the influence of three distinct aspects of the vesicular ensemble (vesicle size, membrane composition, and motor density) on cargo transport. We also demonstrate the presence of vesicle populations that travel at velocities up to twice the unloaded velocity of a single motor. These findings serve to bridge the gap between enhanced vesicular velocities measured in vivo and depressed velocities measured in vitro. Myosin Va is an actin-based molecular motor responsible for transport and positioning of a wide array of intracellular cargoes. Although myosin Va motors have been well characterized at the single-molecule level, physiological transport is carried out by ensembles of motors. Studies that explore the behavior of ensembles of molecular motors have used nonphysiological cargoes such as DNA linkers or glass beads, which do not reproduce one key aspect of vesicular systems--the fluid intermotor coupling of biological lipid membranes. Using a system of defined synthetic lipid vesicles (100- to 650-nm diameter) composed of either 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) (fluid at room temperature) or 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) (gel at room temperature) with a range of surface densities of myosin Va motors (32-125 motors per m2), we demonstrate that the velocity of vesicle transport by ensembles of myosin Va is sensitive to properties of the cargo. Gel-state DPPC vesicles bound with multiple motors travel at velocities equal to or less than vesicles with a single myosin Va ([~]450 nm/s), whereas surprisingly, ensembles of myosin Va are able to transport fluid-state DOPC vesicles at velocities significantly faster (>700 nm/s) than a single motor. To explain these data, we developed a Monte Carlo simulation that suggests that these reductions in velocity can be attributed to two distinct mechanisms of intermotor interference (i.e., load-dependent modulation of stepping kinetics and binding-site exclusion), whereas faster transport velocities are consistent with a model wherein the normal stepping behavior of the myosin is supplemented by the preferential detachment of the trailing motor from the actin track.
