Regulation of myosin V processivity by calcium at the single molecule level.
Calcium can affect myosin V (myoV) function in at least two ways. The full-length molecule, which adopts a folded inhibited conformation in EGTA, becomes extended and active in the presence of calcium. Calcium also dissociates one or more calmodulin molecules from the extended neck. Here we investigated at the single molecule level how calcium regulates the processive run length of full-length myosin V (dFull) and a truncated dimeric construct (dHMM), which cannot adopt the folded conformation. The processivity of dFull and dHMM is tightly controlled by the calcium and calmodulin concentration, with shorter runs occurring at higher calcium concentration. The data indicate that a calcium-dependent dissociation of calmodulin from the neck region of myoV terminates its processive run. dFull showed unexpected processive movement in EGTA, suggesting that a small population of extended, active molecules are in equilibrium with the inhibited, folded form. Single turnover assays showed that the ATPase activity of the folded full-length molecule is inhibited by more than 50-fold compared with the extended molecule. The results imply that activation and termination of the processive runs of myoV can be accomplished by multiple mechanisms.
Lu
Hailong
H
Department of Molecular Physiology and Biophysics, University of Vermont, Burlington, Vermont 05405, USA.
Krementsova
Elena B
EB
Trybus
Kathleen M
KM
D000199:Actins / Q000737:chemistry; D000251:Adenosine Triphosphatases / Q000737:chemistry; D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D002118:Calcium / Q000737:chemistry / Q000378:metabolism*; D002147:Calmodulin / Q000737:chemistry; D002645:Chickens; D019281:Dimerization; D004533:Egtazic Acid / Q000737:chemistry; D007700:Kinetics; D008954:Models, Biological; D024701:Myosin Type V / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary
D000199:Actins; D002147:Calmodulin; D004533:Egtazic Acid; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D024701:Myosin Type V; D002118:Calcium