Hydrolysis of AMPPNP by the motor domain of ncd, a kinesin‐related protein

FEBS Letters

Tanokura, Masaru¹  Morii, Hisayuki²  Shimizu, Takashi²  Suzuki, Yoshikazu¹ 

[1] Biotechnology Research Center, University of Tokyo, Bunkyo, 113 Tokyo, Japan;National Institute of Bioscience and Human-Technology, Higashi, Tsukuba, 305 Ibaraqi, Japan

关键词: Adenylylimidodiphosphate;    Ncd;    Kinesin;    Molecular motor;    31P NMR;    AMPPCP;    adenylylmethylenediphosphonate;    AMPPN;    adenosine 5′-diphosphoramidate;    AMPPNP;    adenylylimidodiphosphate;    EGTA;    ethylene glycol bis(β-aminoethylether)-N;    N;    N′;    N′-tetraacetic acid;    mant-ADP;    2′(3′)-O-(N-methylanthraniloyl)adenosine 5′-diphosphate;    mant-AMPPNP;    2′(3′)-O-(N-methylanthraniloyl)adenylylimidodiphosphate;    Mops;    3-(N-morpholino)propanesulfonic acid;   

DOI  :  10.1016/S0014-5793(97)00472-9

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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 ≈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.

【 授权许可】

Unknown