Inhibition of muscle force by vanadate.
Vanadate (Vi), an analogue of inorganic phosphate (Pi), is known to bind tightly with a long half life to the myosin MgATPase site, producing a complex which inhibits force. Both of these ligands bind to an actin.myosin.ADP state that follows the release of Pi in the enzymatic cycle, and their effects on muscle fibers and proteins in solution provide information on the properties of this state. The inhibition of active force generation began to occur at a [Vi] of 5 microM and was 90% complete at a [Vi] of 1 mM. Hill plots of the inhibition of force by Vi approximated that expected for a simple binding isotherm. Similar plots were obtained at both 25 degrees C and 5 degrees C. A simple binding isotherm is not expected to occur in a muscle fiber where steric constraints imposed by the intact filaments should introduce more complexity into the energetics of ligand binding. The inhibition of MgATPase activity for acto-subfragment-1 to 50% of controls occurred at a [Vi] which was only 20-fold higher than that required to inhibit force generation in fibers to the same level. Some models of actomyosin interactions would predict that the range of [Vi] required for complete force inhibition in fibers and the difference in the [Vi] required for inhibition in fibers and of myosin in solution would both be much larger.
Wilson
G J
GJ
Department of Pathology, University of Sydney, New South Wales, Australia.
Shull
S E
SE
Cooke
R
R
D000199:Actins / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D001667:Binding, Competitive; D055592:Biophysical Phenomena; D001703:Biophysics; D017301:Ca(2+) Mg(2+)-ATPase / Q000037:antagonists & inhibitors / Q000378:metabolism; D002118:Calcium / Q000494:pharmacology; D066298:In Vitro Techniques; D007700:Kinetics; D008024:Ligands; D008954:Models, Biological; D009119:Muscle Contraction / Q000187:drug effects* / Q000502:physiology; D015879:Myosin Subfragments / Q000378:metabolism; D009218:Myosins / Q000037:antagonists & inhibitors; D010710:Phosphates / Q000378:metabolism / Q000494:pharmacology; D016658:Psoas Muscles / Q000187:drug effects / Q000502:physiology; D011817:Rabbits; D013696:Temperature; D014638:Vanadates / Q000378:metabolism / Q000494:pharmacology*
D000199:Actins; D008024:Ligands; D015879:Myosin Subfragments; D010710:Phosphates; D014638:Vanadates; D017301:Ca(2+) Mg(2+)-ATPase; D009218:Myosins; D002118:Calcium