Kinetic mechanism of the fastest motor protein, Chara myosin.
Chara corallina class XI myosin is by far the fastest molecular motor. To investigate the molecular mechanism of this fast movement, we performed a kinetic analysis of a recombinant motor domain of Chara myosin. We estimated the time spent in the strongly bound state with actin by measuring rate constants of ADP dissociation from actin.motor domain complex and ATP-induced dissociation of the motor domain from actin. The rate constant of ADP dissociation from acto-motor domain was >2800 s(-1), and the rate constant of ATP-induced dissociation of the motor domain from actin at physiological ATP concentration was 2200 s(-1). From these data, the time spent in the strongly bound state with actin was estimated to be <0.82 ms. This value is the shortest among known values for various myosins and yields the duty ratio of <0.3 with a V(max) value of the actin-activated ATPase activity of 390 s(-1). The addition of the long neck domain of myosin Va to the Chara motor domain largely increased the velocity of the motility without increasing the ATP hydrolysis cycle rate, consistent with the swinging lever model. In addition, this study reveals some striking kinetic features of Chara myosin that are suited for the fast movement: a dramatic acceleration of ADP release by actin (1000-fold) and extremely fast ATP binding rate.
Ito
Kohji
K
Department of Biology, Chiba University, Inage-ku, Chiba 263-8522, Japan. k-ito@faculty.chiba-u.jp
Ikebe
Mitsuo
M
Kashiyama
Taku
T
Mogami
Toshifumi
T
Kon
Takahide
T
Yamamoto
Keiichi
K
D000199:Actins / Q000737:chemistry* / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000737:chemistry / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000737:chemistry / Q000378:metabolism; D020418:Algal Proteins / Q000737:chemistry* / Q000378:metabolism; D044784:Chara / Q000737:chemistry* / Q000378:metabolism; D006868:Hydrolysis; D007700:Kinetics; D024701:Myosin Type V / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary
D000199:Actins; D020418:Algal Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D024701:Myosin Type V