Purified kinesin promotes vesicle motility and induces active sliding between microtubules in vitro.
We examined the ability of kinesin to support the movement of adrenal medullary chromaffin granules on microtubules in a defined in vitro system. We found that kinesin and ATP are all that is required to support efficient (33% vesicle motility) and rapid (0.4-0.6 micron/s) translocation of secretory granule membranes on microtubules in the presence of a low-salt motility buffer. Kinesin also induced the formation of microtubule asters in this buffer, with the plus ends of microtubules located at the center of each aster. This observation indicates that kinesin is capable of promoting active sliding between microtubules toward their respective plus ends, a movement analogous to that of anaphase b in the mitotic spindle. The fact that vesicle translocation, microtubule sliding, and microtubule-dependent kinesin ATPase activities are all enhanced in low-salt buffer establishes a functional parallel between this translocator and other motility ATPases, myosin, and dynein.
Urrutia
R
R
Laboratory of Cellular Biology, National Institute on Deafness and other Communication Disorders, Bethesda, MD 20892.
McNiven
M A
MA
Albanesi
J P
JP
Murphy
D B
DB
Kachar
B
B
D000251:Adenosine Triphosphatases / Q000302:isolation & purification / Q000494:pharmacology* / Q000502:physiology; D000313:Adrenal Medulla / Q000502:physiology* / Q000648:ultrastructure; D000818:Animals; D001921:Brain / Q000502:physiology; D017301:Ca(2+) Mg(2+)-ATPase / Q000378:metabolism; D002417:Cattle; D002837:Chromaffin Granules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D016547:Kinesins; D007700:Kinetics; D008868:Microtubule Proteins / Q000494:pharmacology*; D008870:Microtubules / Q000187:drug effects / Q000502:physiology* / Q000648:ultrastructure; D009994:Osmolar Concentration
D008868:Microtubule Proteins; D000251:Adenosine Triphosphatases; D017301:Ca(2+) Mg(2+)-ATPase; D016547:Kinesins