Dictyostelium myosin-5b is a conditional processive motor.
Dictyostelium myosin-5b is the gene product of myoJ and one of two closely related myosin-5 isoenzymes produced in Dictyostelium discoideum. Here we report a detailed investigation of the kinetic and functional properties of the protein. In standard assay buffer conditions, Dictyostelium myosin-5b displays high actin affinity in the presence of ADP, fast ATP hydrolysis, and a high steady-state ATPase activity in the presence of actin that is rate limited by ADP release. These properties are typical for a processive motor that can move over long distances along actin filaments without dissociating. Our results show that a physiological decrease in the concentration of free Mg(2+)-ions leads to an increased rate of ADP release and shortening of the fraction of time the motor spends in the strong actin binding states. Consistently, the ability of the motor to efficiently translocate actin filaments at very low surface densities decreases with decreasing concentrations of free Mg(2+)-ions. In addition, we provide evidence that the observed changes in Dd myosin-5b motor activity are of physiological relevance and propose a mechanism by which this molecular motor can switch between processive and non-processive movement.
Taft
Manuel H
MH
Institute for Biophysical Chemistry, OE 4350, Hannover Medical School, Feodor-Lynen-Str. 5, D-30625 Hannover, Germany.
Hartmann
Falk K
FK
Rump
Agrani
A
Keller
Heiko
H
Chizhov
Igor
I
Manstein
Dietmar J
DJ
Tsiavaliaris
Georgios
G
D008841:Actin Cytoskeleton / Q000378:metabolism; D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D002413:Cations, Divalent / Q000378:metabolism; D004023:Dictyostelium / Q000201:enzymology*; D007527:Isoenzymes / Q000378:metabolism; D008274:Magnesium / Q000378:metabolism; D009218:Myosins / Q000378:metabolism*; D015800:Protozoan Proteins / Q000378:metabolism*
D002413:Cations, Divalent; D007527:Isoenzymes; D015800:Protozoan Proteins; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D009218:Myosins; D008274:Magnesium