Competitive action of divalent cations and D600 in frog slow muscle fibers

Single, slow muscle fibers from Rana temporaria were equilibrated in normal Ringer's. 95 mmol/liter K(+)-solution containing various concentrations of Ca2+, Ni2+, Mn2+ or Mg2+ was applied, and the ensuing contractures were recorded isometrically. While peak tension (Fmax) was little affected, maintained tension (measured 1 min after onset of contracture) strongly depended on the concentration and species of divalent cations. Tension was maintained at its peak value in the presence of all species of divalent cations provided their concentrations were adequately increased. Dose-response curves were hyperbolic; Lineweaver-Burk plots revealed straight lines with different slopes intersecting near 1/Fmax, and indicating the following order of efficiency: Ni2+ greater than Ca2+ greater than Mn2+ much greater than Mg2+. Hill plots for these cations resulted in straight lines with slopes near 1. Qualitatively similar relationships were obtained with contracture solutions containing D600 (3-12 mumol/liter). However, under these conditions higher concentrations of Ca2+ or Ni2+ were required in order to fully maintain tension. After a step concentration change in the medium during contracture, the effects of Ca2+ or D600 were detectable only after a delay of 9 and 18 sec, respectively. It is concluded that divalent cations and D600 compete for the same binding site according to a 1:1 reaction. This site is presumably located inside the transverse tubular system and controls inactivation of the contractile force.