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- Title
Biochemical characterization of the novel rice kinesin K23 and its kinetic study using fluorescence resonance energy transfer between an intrinsic tryptophan residue and a fluorescent ATP analogue.
- Authors
Umezu, Nozomi; Hanzawa, Nobue; Yamada, Masafumi D.; Kondo, Kazunori; Mitsui, Toshiaki; Maruta, Shinsaku
- Abstract
We previously demonstrated that the rice kinesin K16, which belongs to the kinesin-7 subfamily, has unique enzymatic properties and atomic structure within key functional regions. In this study, we focused on a novel rice plant kinesin, K23, which also belongs to the kinesin-7 subfamily. The biochemical characterization of the K23 motor domain (K23MD) was studied and compared with the rice kinesin K16 and other related kinesins. K23 exhibits ∼45-fold (1.3 Pi mol−1site mol−1s−1) lower microtubule-dependent ATPase activity than conventional kinesins, whereas its affinity for microtubules is comparable with conventional kinesins. MgADP-free K23 is unstable compared with the unusually stable MgADP-free K16MD. The enzymatic properties of K23MD are somewhat different from those of K16. We used a fluorescent ATP analogue 2′(3′)-O-(N′-methylanthraniloyl)-ATP (mant-ATP) for the kinetic characterization of K23. The fluorescence of mant-ATP was not significantly altered during its hydrolysis by K23. However, significant fluorescence resonance energy transfer (FRET) between mant-ATP and W21 in the motor domain was observed. The kinetic study using FRET revealed that K23 has unique kinetic characteristics when compared with other kinesins.
- Subjects
KINESIN; FLUORESCENCE; ENERGY transfer; TRYPTOPHAN; ADENOSINE triphosphate
- Publication
Journal of Biochemistry, 2011, Vol 149, Issue 5, p539
- ISSN
0021-924X
- Publication type
Article
- DOI
10.1093/jb/mvr012