Dynamics of myo1c (myosin-ibeta ) lipid binding and dissociation.
Myosin-I is the single-headed member of the myosin superfamily that associates with lipid membranes. Biochemical experiments have shown that myosin-I membrane binding is the result of electrostatic interactions between the basic tail domain and acidic phospholipids. To better understand the dynamics of myosin-I membrane association, we measured the rates of association and dissociation of a recombinant myo1c tail domain (which includes three IQ domains and bound calmodulins) to and from large unilamellar vesicles using fluorescence resonance energy transfer. The apparent second-order rate constant for lipid-tail association in the absence of calcium is fast with nearly every lipid-tail collision resulting in binding. The rate of binding is decreased in the presence of calcium. Time courses of myo1c-tail dissociation are best fit by two exponential rates: a fast component that has a rate that depends on the ratio of acidic phospholipid to myo1c-tail (phosphatidylserine (PS)/tail) and a slow component that predominates at high PS/tail ratios. The dissociation rate of the slow component is slower than the myo1c ATPase rate, suggesting that myo1c is able to stay associated with the lipid membrane during multiple catalytic cycles of the motor. Calcium significantly increases the lifetimes of the membrane-bound state, resulting in dissociation rates 0.001 s(-1).
Tang
Nanyun
N
Pennsylvania Muscle Institute and the Department of Physiology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104-6085, USA.
Lin
Tianming
T
Ostap
E Michael
EM
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D003001:Cloning, Molecular; D004229:Dithiothreitol; D007700:Kinetics; D050356:Lipid Metabolism; D008081:Liposomes; D024461:Myosin Type I / Q000737:chemistry* / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry / Q000378:metabolism; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D055672:Static Electricity; D014714:Vertebrates
D008081:Liposomes; D010743:Phospholipids; D011994:Recombinant Proteins; D024461:Myosin Type I; D002118:Calcium; D004229:Dithiothreitol