Hydrolysis of AMPPNP by the motor domain of ncd, a kinesin-related protein.
AMPPNP was found to be hydrolyzed by the motor domain of ncd (the product of a Drosophila gene, non-claret disjunctional), a kinesin-related protein. This hydrolysis could be monitored by 31P NMR spectroscopy and by an assay of phosphate, one of the products of the hydrolysis. The rate was approximately 0.00004 s(-1), 1% of the ATP turnover rate. The AMPPNP turnover was not stimulated by microtubules. Kinesin motor domain also turned over AMPPNP but at a somewhat lower rate. Although the turnover was slow, the present finding may present an important caveat, since AMPPNP has been widely used for investigations of kinesin and kinesin-related proteins as a non-hydrolyzable ATP analogue.
Suzuki
Y
Y
Biotechnology Research Center, University of Tokyo, Bunkyo, Japan.
Shimizu
T
T
Morii
H
H
Tanokura
M
M
D000255:Adenosine Triphosphate / Q000037:antagonists & inhibitors / Q000378:metabolism; D000266:Adenylyl Imidodiphosphate / Q000378:metabolism* / Q000494:pharmacology; D000818:Animals; D002942:Circular Dichroism; D029721:Drosophila Proteins; D004331:Drosophila melanogaster; D006801:Humans; D006868:Hydrolysis; D016547:Kinesins / Q000737:chemistry / Q000494:pharmacology*; D009682:Magnetic Resonance Spectroscopy; D008868:Microtubule Proteins / Q000737:chemistry / Q000494:pharmacology*; D008870:Microtubules / Q000502:physiology; D017434:Protein Structure, Tertiary
D029721:Drosophila Proteins; D008868:Microtubule Proteins; C066696:ncd protein, Drosophila; D000266:Adenylyl Imidodiphosphate; D000255:Adenosine Triphosphate; D016547:Kinesins