Temperature dependent properties of a kinesin-3 motor protein from Thermomyces lanuginosus.
Kinesins are cytoskeletal motor proteins that share a common mechanochemical motor domain, and are responsible for trafficking macromolecules. Here we report the cloning and characterization of a monomeric, kinesin-3 (TKIN) from Thermomyces lanuginosus. TKIN displayed a maximum rate of ATP hydrolysis at approximately 55 degrees C; the K(m)(ATP) was also significantly greater at 50 degrees C. Gliding motility rates reached a maximum of 5.5 microms(-1) at 45 degrees C, which is among the highest rates reported for kinesin. Arrhenius energy barriers were calculated to be approximately 103 kJmol(-1), nearly twofold greater than other mesophilic kinesin motors. The enthalpy of activation and entropy activation of TKIN were also significantly greater when compared to other mesophilic kinesins. A thermally induced aggregation of TKIN, which could be moderated by the addition of ATP, was observed at temperatures above 45 degrees C. Together, these results illustrate the kinetic response and stability of this unique motor protein at elevated temperatures.
Rivera
Susan B
SB
Biomolecular Interfaces and Systems Department, Sandia National Laboratories, P.O. Box 5800, MS-1413, Albuquerque, NM 87185-1413, USA.
Koch
Steven J
SJ
Bauer
Joseph M
JM
Edwards
J Matthew
JM
Bachand
George D
GD
D000251:Adenosine Triphosphatases / Q000378:metabolism; D001203:Ascomycota / Q000235:genetics / Q000502:physiology*; D006358:Hot Temperature; D016547:Kinesins / Q000378:metabolism*; D007700:Kinetics; D008870:Microtubules / Q000378:metabolism*; D020409:Molecular Motor Proteins / Q000378:metabolism*; D013696:Temperature
D020409:Molecular Motor Proteins; D000251:Adenosine Triphosphatases; D016547:Kinesins