Abstract
We have investigated the structural changes that occur in the molecular motor kinesin during its ATPase cycle, utilizing two bacterially expressed constructs. The structure of both constructs has been examined as a function of the nature of the nucleotide intermediate occupying the active site by means of sedimentation velocity, sedimentation equilibrium, fluorescence solute quenching, fluorescence anisotropy decay, and limited proteolysis. While the molecular weight of monomeric and dimeric human kinesin constructs, as measured by sedimentation velocity and sedimentation equilibrium, and the tryptic cleavage pattern are unaffected by the nucleotide intermediate occupying the active site, significant changes in the rotational correlation time of fluorescently labeled kinesin-nucleotide intermediates can be detected. These results suggest that kinesin contains an internal "hinge" whose flexibility varies through the course of the ATPase cycle. In prehydrolytic, "strong" binding states, this hinge is relatively rigid, while in posthydrolytic, "weak" binding states, it is more flexible. Our results, in conjunction with anisotropy decay studies of myosin, suggest that these two molecular motors may share a common structural feature...Continue Reading
References
Jun 1, 1992·Analytical Biochemistry·W F Stafford
Jan 1, 1992·Annual Review of Cell Biology·S A Endow, M A Titus
May 22, 1992·Biochimica Et Biophysica Acta·C K WangH C Cheung
Jun 1, 1992·The Journal of Cell Biology·F NavoneR D Vale
Apr 3, 1990·Biochemistry·C R CremoR G Yount
Nov 9, 1989·Nature·J HowardR D Vale
Nov 1, 1989·Analytical Biochemistry·S C Gill, P H von Hippel
Sep 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·D D Hackney
Jan 1, 1985·Methods in Enzymology·G C Na, S N Timasheff
Mar 31, 1981·Biochemistry·J W Shriver, B D Sykes
Oct 27, 1981·Biochemistry·J W Shriver, B D Sykes
Dec 1, 1981·Biophysical Journal·M L JohnsonH R Halvorson
Jul 20, 1995·Nature·K HiroseL A Amos
Apr 11, 1995·Biochemistry·J J CorreiaK A Johnson
Mar 1, 1995·Biophysical Journal·J Ma
Jan 1, 1994·Methods in Enzymology·W F Stafford
Feb 23, 1995·Nature·S P GilbertK A Johnson
Feb 22, 1994·Biochemistry·S P Gilbert, K A Johnson
Jul 2, 1993·Science·I RaymentH M Holden
Jan 1, 1993·Annual Review of Biochemistry·R A Walker, M P Sheetz
Jan 14, 1993·Nature·L Romberg, R D Vale
Jan 18, 1996·Nature·A S KashinaJ M Scholey
Apr 11, 1996·Nature·F J KullR D Vale
Apr 11, 1996·Nature·E P SablinR J Fletterick
Apr 19, 1996·The Journal of Biological Chemistry·S S RosenfeldH C Cheung
Apr 16, 1996·Biophysical Chemistry·B C PhanE Reisler
Citations
Jul 24, 1998·Biophysical Journal·W Wriggers, K Schulten
Mar 8, 2011·Journal of Molecular Biology·P Keith VeroneseAaron L Lucius
Nov 22, 2014·ELife·Zhiguo ShangCharles V Sindelar
Oct 6, 2000·The Journal of Biological Chemistry·F KozielskiM H Koch
Jul 21, 2005·The Journal of Biological Chemistry·Peter HöökRichard B Vallee
Apr 16, 1998·The Journal of Biological Chemistry·H L SweeneyJ R Sellers
Oct 24, 1998·The Journal of Biological Chemistry·S S RosenfeldH L Sweeney
Dec 18, 1998·The Journal of Biological Chemistry·K A FosterS P Gilbert
Jul 18, 2002·The Journal of Biological Chemistry·Steven S RosenfeldPeter H King
Jan 26, 2006·The Journal of Biological Chemistry·Zoltan MaligaSteven S Rosenfeld
May 29, 2000·The Journal of Biological Chemistry·S S RosenfeldH L Sweeney
Jun 15, 2000·The Journal of Biological Chemistry·J XingS S Rosenfeld
Nov 21, 1997·Journal of Molecular Biology·I M CrevelR A Cross
Mar 2, 1999·Journal of Structural Biology·A Lockhart, J Kendrick-Jones
Jan 3, 2001·Methods : a Companion to Methods in Enzymology·S P Gilbert, A T Mackey
Feb 28, 2002·Current Biology : CB·Junichiro YajimaYoko Y Toyoshima
Jul 25, 1997·Cell·G WoehlkeR D Vale