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Biophysical Journal 84:1833-1843 (2003)
© 2003 The Biophysical Society

Single Fungal Kinesin Motor Molecules Move Processively along Microtubules

Stefan Lakämper*,{dagger},{ddagger}, Athina Kallipolitou{dagger}, Günther Woehlke{dagger}, Manfred Schliwa{dagger} and Edgar Meyhöfer*,{ddagger}

* Cellular and Molecular Physiology, Medical School Hannover, 30635 Hannover, Germany; {dagger} Adolf-Butenandt-Institute of Cell Biology, Ludwig-Maximilians-University Munich, 80336 Munich, Germany; and {ddagger} Mechanical Engineering, University of Michigan, Ann Arbor, Michigan 48109 USA

Correspondence: Address reprint requests to Edgar Meyhöfer, E-mail: meyhofer{at}umich.edu.

Conventional kinesins are two-headed molecular motors that move as single molecules micrometer-long distances on microtubules by using energy derived from ATP hydrolysis. The presence of two heads is a prerequisite for this processive motility, but other interacting domains, like the neck and K-loop, influence the processivity and are implicated in allowing some single-headed kinesins to move processively. Neurospora kinesin (NKin) is a phylogenetically distant, dimeric kinesin from Neurospora crassa with high gliding speed and an unusual neck domain. We quantified the processivity of NKin and compared it to human kinesin, HKin, using gliding and fluorescence-based processivity assays. Our data show that NKin is a processive motor. Single NKin molecules translocated microtubules in gliding assays on average 2.14 µm (N = 46). When we tracked single, fluorescently labeled NKin motors, they moved on average 1.75 µm (N = 182) before detaching from the microtubule, whereas HKin motors moved shorter distances (0.83 µm, N = 229) under identical conditions. NKin is therefore at least twice as processive as HKin. These studies, together with biochemical work, provide a basis for experiments to dissect the molecular mechanisms of processive movement.




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