Cytoplasmic dynein is a vesicle protein.
Microtubule-based organelle transport is thought to be mediated by the force-generating proteins cytoplasmic dynein and kinesin. These motor proteins have been characterized based on their ability to associate with and translocate microtubules. We show here that cytoplasmic dynein is also present as a peripheral membrane protein of purified synaptic vesicles. The vesicle-associated cytoplasmic dynein is identified by its photo-induced cleavage in the presence of ATP and vanadate. Purified, soluble cytoplasmic dynein is competent to bind to vesicle membranes stripped of endogenous peripheral membrane proteins by alkaline pH. Dynein binding to membranes is saturable at a concentration of 1.00 +/- 0.15 pmol/micrograms vesicle protein and has a dissociation constant of 22.3 +/- 2.4 nM. The association of cytoplasmic dynein with the membrane cannot be reversed by incubation with ATP. Furthermore, following binding to membranes, dynein retains its ability to bind ATP and to be photo-cleaved in the presence of vanadate. The presence of cytoplasmic dynein on synaptic vesicles and its ability to bind to extracted membranes supports current models of microtubule-based organelle translocation.
Lacey
M L
ML
Department of Biology, University of California, Riverside 92521.
Haimo
L T
LT
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D002850:Chromatography, Gel; D003593:Cytoplasm / Q000378:metabolism*; D004398:Dyneins / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D016547:Kinesins / Q000378:metabolism; D007700:Kinetics; D008869:Microtubule-Associated Proteins / Q000378:metabolism; D008970:Molecular Weight; D013574:Synaptosomes / Q000378:metabolism; D014638:Vanadates / Q000378:metabolism
D008869:Microtubule-Associated Proteins; D014638:Vanadates; D000255:Adenosine Triphosphate; D004398:Dyneins; D016547:Kinesins