Substrate-concentration of dependences of tension, shortening velocity and ATPase activity of glycerinated single muscle fibers.
Tension P0 and ATPase activity J0 of glycerinated single muscle fibers under isometric concentration as well as the velocity V0 of unloaded shortening were measured as a function of substrate concentration [S]. The stiffness of fibers with sinusoidal length changes at 1 kHz was used as a qualitative measure of the amount of rigor complex. P0 is an increasing function of [S] at low substrate concentrations and has a broad maximum at about 10-40 micrometers MgATP. In this concentration range, 10-40 micrometers, V0 still has a very small value. Then it increases and finally reaches at a plateau at about 1 mM MgATP. J0 increases as P0 does. However, it reaches at a saturated level at about the same concentration as V0. Either 0.5 mM 8-BrATP or 1 mM PPi was added to the substrate solutions to reduce the amount of rigor complex at low substrate concentrations. The addition of PPi of 8-BrATP decreases P0 dominantly at low concentrations of substrate and shifts the maximum to about 100 micrometers MgATP. 8-BrATP considerably increases V0 at low substrate concentrations while V0 is decreased by added PPi. The temperature coefficients, Q10 values were obtained for P0, J0, and V0. The values are essentially constant, 2.1-2.4, in the cases of P0 and J0, and about the same values were found for V0 at very low substrate concentrations. However, they become about 3.3 in the concentration range from 34 micrometers and 2.3 mM. The P-V relation was obtained at 11 micrometers and 2.3 micrometers MgATP. The normalized P-V relation at 11 micrometers was unchanged when 8-BrATP was added. The results are discussed in connection with the mechanism of actomyosin-ATPase activity as well as that of the elementary cycle of the motive force generation.
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D011756:Diphosphates / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D009119:Muscle Contraction; D009218:Myosins / Q000378:metabolism*; D011817:Rabbits
D011756:Diphosphates; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D009218:Myosins