title: Kinesin's step dissected with single-motor FRET.
abstract: The motor protein Kinesin-1 drives intracellular transport along microtubules, with each of its two motor domains taking 16-nm steps in a hand-over-hand fashion. The way in which a single-motor domain moves during a step is unknown. Here, we use Förster resonance energy transfer (FRET) between fluorescent labels on both motor domains of a single kinesin. This approach allows us to resolve the relative distance between the motor domains and their relative orientation, on the submillisecond timescale, during processive stepping. We observe transitions between high and low FRET values for certain kinesin constructs, depending on the location of the labels. These results reveal that, during a step, a kinesin motor domain dwells in a well-defined intermediate position for approximately 3 ms.
authors.affiliation: Department of Physics and Astronomy and Laser Centre, VU University, de Boelelaan 1081, 1081 HV, Amsterdam, The Netherlands.
authors.forename: Sander
authors.initials: S
authors.lastname: Verbrugge
authors.forename: Zdenek
authors.initials: Z
authors.lastname: Lansky
authors.forename: Erwin J G
authors.initials: EJ
authors.lastname: Peterman
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D019943:Amino Acid Substitution; D031541:Fluorescence Resonance Energy Transfer; D006801:Humans; D066298:In Vitro Techniques; D016547:Kinesins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism; D008954:Models, Biological; D008958:Models, Molecular; D020409:Molecular Motor Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D009010:Monte Carlo Method; D016297:Mutagenesis, Site-Directed; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D020409:Molecular Motor Proteins; D011994:Recombinant Proteins; D000255:Adenosine Triphosphate; D016547:Kinesins