Kinesin's second step.

We have identified dimeric kinesin mutants that become stalled on the microtubule after one ATP turnover, unable to bind and hydrolyze ATP at their second site. We have used these mutants to determine the regulatory signal that allows ATP to bind to the forward head, such that processive movement can continue. The results show that phosphate release occurs from the rearward head before detachment, and detachment triggers active-site accessibility for ATP binding at the forward head. This mechanism, in which the rearward head controls the behavior of the forward head, may be conserved among processive motors.

[1]  P. Kollman,et al.  Closing of the Nucleotide Pocket of Kinesin-Family Motors upon Binding to Microtubules , 2003, Science.

[2]  Hernando Sosa,et al.  Configuration of the two kinesin motor domains during ATP hydrolysis , 2003, Nature Structural Biology.

[3]  S. Rosenfeld,et al.  ATP Reorients the Neck Linker of Kinesin in Two Sequential Steps* , 2001, The Journal of Biological Chemistry.

[4]  Christopher M. Farrell,et al.  A Kinesin Switch I Arginine to Lysine Mutation Rescues Microtubule Function* , 2003, Journal of Biological Chemistry.

[5]  M. Schnitzer,et al.  Force production by single kinesin motors , 2000, Nature Cell Biology.

[6]  J. Corrie,et al.  Direct, real-time measurement of rapid inorganic phosphate release using a novel fluorescent probe and its application to actomyosin subfragment 1 ATPase. , 1994, Biochemistry.

[7]  D. Hackney,et al.  Evidence for alternating head catalysis by kinesin during microtubule-stimulated ATP hydrolysis. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[8]  S. Rosenfeld,et al.  Measuring Kinesin's First Step* , 2002, The Journal of Biological Chemistry.

[9]  E. Mandelkow,et al.  Nucleotide‐induced conformations in the neck region of dimeric kinesin , 2003, The EMBO journal.

[10]  Christopher M. Farrell,et al.  The Role of ATP Hydrolysis for Kinesin Processivity* , 2002, The Journal of Biological Chemistry.

[11]  H. Sweeney,et al.  Kinetic Mechanism and Regulation of Myosin VI* , 2001, The Journal of Biological Chemistry.

[12]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[13]  E. Taylor,et al.  Mechanism of microtubule kinesin ATPase. , 1995, Biochemistry.

[14]  K. Johnson,et al.  Pathway of ATP hydrolysis by monomeric and dimeric kinesin. , 1998, Biochemistry.

[15]  Justin E. Molloy,et al.  The gated gait of the processive molecular motor, myosin V , 2002, Nature Cell Biology.

[16]  Polly M. Fordyce,et al.  Stepping and Stretching , 2003, The Journal of Biological Chemistry.

[17]  J. Gelles,et al.  Distinguishing Inchworm and Hand-Over-Hand Processive Kinesin Movement by Neck Rotation Measurements , 2002, Science.

[18]  Roger Cooke,et al.  Two conformations in the human kinesin power stroke defined by X-ray crystallography and EPR spectroscopy , 2002, Nature Structural Biology.

[19]  E. Mandelkow,et al.  X-ray structure of motor and neck domains from rat brain kinesin. , 1997, Biochemistry.

[20]  D. Rose,et al.  Lethal Kinesin Mutations Reveal Amino Acids Important for ATPase Activation and Structural Coupling* , 1999, The Journal of Biological Chemistry.

[21]  E. Taylor,et al.  Interacting Head Mechanism of Microtubule-Kinesin ATPase* , 1997, The Journal of Biological Chemistry.

[22]  W. Schief,et al.  Conformational changes during kinesin motility. , 2001, Current opinion in cell biology.

[23]  W. Wriggers,et al.  Kinesin Has Three Nucleotide-dependent Conformations , 2000, The Journal of Biological Chemistry.

[24]  K. Johnson,et al.  Pre-steady-state kinetics of the microtubule-kinesin ATPase. , 1994, Biochemistry.

[25]  Roger Cooke,et al.  A structural change in the kinesin motor protein that drives motility , 1999, Nature.

[26]  Manfred Schliwa,et al.  Molecular motors , 2003, Nature.

[27]  Kenneth H. Downing,et al.  Structure of the αβ tubulin dimer by electron crystallography , 1998, Nature.

[28]  R Vale,et al.  Thermodynamic properties of the kinesin neck-region docking to the catalytic core. , 2003, Biophysical journal.

[29]  R. Vale,et al.  The way things move: looking under the hood of molecular motor proteins. , 2000, Science.

[30]  Matthew J Tyska,et al.  Myosin-V motility: these levers were made for walking. , 2003, Trends in cell biology.

[31]  A. Hoenger,et al.  Motor domain mutation traps kinesin as a microtubule rigor complex. , 2003, Biochemistry.

[32]  David D Hackney Pathway of ADP-stimulated ADP release and dissociation of tethered kinesin from microtubules. Implications for the extent of processivity. , 2002, Biochemistry.

[33]  E. Mandelkow,et al.  The structural and mechanochemical cycle of kinesin. , 1998, Trends in biochemical sciences.

[34]  K. Johnson,et al.  Alternating site mechanism of the kinesin ATPase. , 1998, Biochemistry.

[35]  Susan P. Gilbert,et al.  Pathway of processive ATP hydrolysis by kinesin , 1995, Nature.