Cooperativity of actin-activated ATPase of gizzard heavy meromyosin in the presence of gizzard tropomyosin.
The mechanism for the potentiation of the actin-activated ATPase of smooth muscle myosin by tropomyosin is investigated using smooth muscle actin, tropomyosin, and heavy meromyosin. In the presence of tropomyosin, an increase in Vmax occurs with no effect on KATPase and Kbinding at 20 mM ionic strength. Utilizing N-ethylmaleimide-treated subfragment-1, which forms rigor complexes with actin in the presence of ATP but does not have ATPase activity, experiments were carried out to determine if the tropomyosin-actin complex exists in both the turned-off and turned-on forms as in the skeletal muscle system. At both 60 and 100 mM ionic strengths, the presence of rigor complexes on the smooth muscle actin filament containing bound tropomyosin causes a 2-3-fold increase in Vmax and about a 3-fold increase in KATPase, resulting in about a 4-fold increase in ATPase activity at moderate actin concentration. The increase in KATPase is correlated with an increase in Kbinding. The finding that rigor complexes increase Vmax and the binding constant for heavy meromyosin to tropomyosin-actin at an ionic strength close to physiological conditions indicates that the tropomyosin-actin complex can be turned on by rigor complexes in a cooperative manner. However, in contrast to the situation in the skeletal muscle system, the increase in KATPase is associated with a corresponding increase in Kbinding. Furthermore, there is only a 3-fold increase in KATPase in the smooth muscle system rather than a 10-fold increase as in the skeletal muscle system.
Chacko
S
S
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia 19104.
Eisenberg
E
E
D000199:Actins / Q000378:metabolism*; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D002918:Chymotrypsin; D004789:Enzyme Activation; D000070559:Gizzard, Non-avian / Q000378:metabolism; D007700:Kinetics; D009119:Muscle Contraction; D009130:Muscle, Smooth / Q000378:metabolism*; D015879:Myosin Subfragments / Q000378:metabolism*; D009994:Osmolar Concentration; D010449:Peptide Mapping; D014335:Tropomyosin / Q000378:metabolism*
D000199:Actins; D015879:Myosin Subfragments; D014335:Tropomyosin; D002918:Chymotrypsin; D000251:Adenosine Triphosphatases