title: Origins of reversed directionality in the ncd molecular motor.
abstract: The head or motor domain of the ncd (non-claret disjunctional) molecular motor is 41% identical to that of kinesin, yet moves along microtubules in the opposite direction to kinesin. We show here that despite the reversed directionality of ncd, its kinetics in solution are homologous in key respects to those of kinesin. The rate limiting step, ADP release, occurs at 0.0033 s-1 at 100 mM NaCl and is accelerated approximately 1000-fold when the motor binds to microtubules. Other reaction steps are all very fast (> 0.1 s-1) compared with ADP release, and the motor is consequently paused in the ncd.ADP state until microtubule binding occurs (Kd = 2 microM), at which point ADP release is triggered and the motor locks onto the microtubule in a rigor-like state. These data identify close functional homology between the strong binding states of kinesin and ncd, and in view of this we discuss a possible mechanism for directional reversal, in which the strong binding states of ncd and kinesin are functionally identical, but the weak binding states are biased in opposite directions.
authors.affiliation: Molecular Motors Group, Marie Curie Research Institute, Surrey, UK.
authors.forename: A
authors.initials: A
authors.lastname: Lockhart
authors.forename: R A
authors.initials: RA
authors.lastname: Cross
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism; D002478:Cells, Cultured; D029721:Drosophila Proteins; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism; D016547:Kinesins / Q000378:metabolism; D007700:Kinetics; D008868:Microtubule Proteins / Q000235:genetics / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D009711:Nucleotides / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D029721:Drosophila Proteins; D008868:Microtubule Proteins; D009711:Nucleotides; D011993:Recombinant Fusion Proteins; C066696:ncd protein, Drosophila; D000244:Adenosine Diphosphate; D005982:Glutathione Transferase; D000251:Adenosine Triphosphatases; D016547:Kinesins