Mechanical Properties of a Single-Headed Processive Motor Inner-Arm Dynein Subspecies-c of ChlamydomonasStudied at the Single Molecule Level

摘要

Dynein from inner arms of Chlamydomonasflagella contains sevendistinct subspecies, a through g. Several lines of evidence suggest thesesubspecies play important roles in generating flagellar beating and thatthe different subspecies are functionally diverse. To evaluate theirroles and diversity, the mechanical properties of subspecies-c, which isa single-headed motor, were examined using optical trap nanometry. Apolystyrene bead coated with a small number of subspecies-c moleculeswas captured with the optical trap and brought into contact with amicrotubule fixed to a coverslip. Movements of the bead were measured bya quadrant photodiode sensor with sub-nanometer- and millisecond-resolution.Beads carrying a single active subspecies-c molecule moved processivelyalong the microtubules in 8-nm steps but slipped backwards under highloads. Force-velocity relationships of single subspecies-c molecules werealmost linear and the shapes of the normalized curves at 5 μM and 100μM ATP were similar. These results indicate that dynein subspecies-cfunctions in a very different way from conventional motor proteins, suchas myosin and kinesin, and has properties that could produceself-oscillation in vivo.