title: Two distinct modes of processive kinesin movement in mixtures of ATP and AMP-PNP.
abstract: An enzyme is frequently conceived of as having a single functional mechanism. This is particularly true for motor enzymes, where the necessity for tight coupling of mechanical and chemical cycles imposes rigid constraints on the reaction pathway. In mixtures of substrate (ATP) and an inhibitor (adenosine 5'-(beta,gamma-imido)triphosphate or AMP-PNP), single kinesin molecules move on microtubules in two distinct types of multiple-turnover "runs" that differ in their susceptibility to inhibition. Longer (less susceptible) runs are consistent with movement driven by the alternating-sites mechanism previously proposed for uninhibited kinesin. In contrast, kinesin molecules in shorter runs step with AMP-PNP continuously bound to one of the two active sites of the enzyme. Thus, in this mixture of substrate and inhibitor, kinesin can function as a motor enzyme using either of two distinct mechanisms. In one of these, the enzyme can accomplish high-duty-ratio processive movement without alternating-sites ATP hydrolysis.
authors.affiliation: Department of Biochemistry, Brandeis University, Waltham, MA 02454, USA.
authors.forename: Radhika
authors.initials: R
authors.lastname: Subramanian
authors.forename: Jeff
authors.initials: J
authors.lastname: Gelles
mesh_terms: D000249:Adenosine Monophosphate / Q000737:chemistry*; D000255:Adenosine Triphosphate / Q000737:chemistry; D000266:Adenylyl Imidodiphosphate / Q000737:chemistry*; D000818:Animals; D001665:Binding Sites; D020134:Catalytic Domain; D004330:Drosophila; D004926:Escherichia coli / Q000378:metabolism; D006868:Hydrolysis; D016547:Kinesins / Q000737:chemistry* / Q000502:physiology*; D007700:Kinetics; D008954:Models, Biological; D015233:Models, Statistical; D009068:Movement; D011487:Protein Conformation; D013997:Time Factors
chemical_list: D000266:Adenylyl Imidodiphosphate; D000249:Adenosine Monophosphate; D000255:Adenosine Triphosphate; D016547:Kinesins