Mechanochemical coupling of two substeps in a single myosin V motor.
Myosin V is a double-headed processive molecular motor that moves along an actin filament by taking 36-nm steps. Using optical trapping nanometry with high spatiotemporal resolution, we discovered that there are two possible pathways for the 36-nm steps, one with 12- and 24-nm substeps, in this order, and the other without substeps. Based on the analyses of effects of ATP, ADP and 2,3-butanedione 2-monoxime (a reagent shown here to slow ADP release from actomyosin V) on the dwell time and the occurrence frequency of the main and the intermediate states, we propose that the 12-nm substep occurs after ATP binding to the bound trailing head and the 24-nm substep results from a mechanical step following the isomerization of an actomyosin-ADP state on the bound leading head. When the isomerization precedes the 12-nm substep, the 36-nm step occurs without substeps.
Uemura
Sotaro
S
Department of Physics, School of Science and Engineering, Waseda University, 3-4-1 Okubo, Shinjuku-ku, Tokyo 169-8555, Japan.
Higuchi
Hideo
H
Olivares
Adrian O
AO
De La Cruz
Enrique M
EM
Ishiwata
Shin'ichi
S
D000199:Actins / Q000737:chemistry; D000205:Actomyosin / Q000737:chemistry; D000244:Adenosine Diphosphate / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D001921:Brain / Q000378:metabolism; D002138:Calibration; D002645:Chickens; D003931:Diacetyl / Q000031:analogs & derivatives* / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D007700:Kinetics; D008954:Models, Biological; D020409:Molecular Motor Proteins / Q000737:chemistry*; D018482:Muscle, Skeletal / Q000378:metabolism; D024701:Myosin Type V / Q000737:chemistry*; D036103:Nanotechnology; D011312:Pressure; D011487:Protein Conformation; D011817:Rabbits; D013997:Time Factors
D000199:Actins; D020409:Molecular Motor Proteins; C004717:diacetylmonoxime; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000205:Actomyosin; D000251:Adenosine Triphosphatases; D024701:Myosin Type V; D003931:Diacetyl