TY - JOUR
T1 - ATP Reorients the Neck Linker of Kinesin in Two Sequential Steps
AU - Rosenfeld, Steven S.
AU - Jefferson, Geraldine M.
AU - King, Peter H.
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2001/10/26
Y1 - 2001/10/26
N2 - Recent models of the kinesin mechanochemical cycle provide some conflicting information on how the neck linker contributes to movement. Some spectroscopic approaches suggest a nucleotide-induced order-to-disorder transition in the neck linker. However, cryoelectron microscopic imaging suggests instead that nucleotide alters the orientation of the neck linker when docked on the microtubule surface. Furthermore, since these studies utilized transition state or non-hydrolyzable nucleotide analogs, it is not clear at what point in the ATPase cycle this reorientation of the neck linker occurs. We have addressed this issue by developing a strategy to examine the effect of nucleotide on the orientation of the neck linker based on the technique of fluorescence resonance energy transfer. Transient kinetic studies utilizing this approach support a model in which ATP binding leads to two sequential isomerizations, the second of which reorients the neck linker in relation to the microtubule surface.
AB - Recent models of the kinesin mechanochemical cycle provide some conflicting information on how the neck linker contributes to movement. Some spectroscopic approaches suggest a nucleotide-induced order-to-disorder transition in the neck linker. However, cryoelectron microscopic imaging suggests instead that nucleotide alters the orientation of the neck linker when docked on the microtubule surface. Furthermore, since these studies utilized transition state or non-hydrolyzable nucleotide analogs, it is not clear at what point in the ATPase cycle this reorientation of the neck linker occurs. We have addressed this issue by developing a strategy to examine the effect of nucleotide on the orientation of the neck linker based on the technique of fluorescence resonance energy transfer. Transient kinetic studies utilizing this approach support a model in which ATP binding leads to two sequential isomerizations, the second of which reorients the neck linker in relation to the microtubule surface.
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U2 - 10.1074/jbc.M103899200
DO - 10.1074/jbc.M103899200
M3 - Article
C2 - 11509561
AN - SCOPUS:0035955690
SN - 0021-9258
VL - 276
SP - 40167
EP - 40174
JO - Journal of Biological Chemistry
JF - Journal of Biological Chemistry
IS - 43
ER -