Effect of various anions on the stability of the coiled coil of skeletal muscle myosin.
The stability of skeletal myosin rod was studied by following the dependence of both papain digestion kinetics and helix-coil transition temperatures on the concentration of neutral salts. The rate of papain-catalyzed digestion of rod to form subfragment 2 and light meromyosin was strongly dependent on chloride concentration but essentially independent of acetate concentration up to 2.0 M. The rod exhibited a biphasic melting curve in 0.6 M NaCl, 5 mM phosphate, and 0.1 mM ethylenediaminetetraacetic acid (EDTA), pH 7.3, with transitions at 45 and 53 degrees C. In 0.6 M CH3COONa, 5 mM phosphate, and 0.1 mM EDTA, pH 7.3, the transitions occurred at 50 and 58 degrees C, respectively. Transition temperatures were obtained with a novel curve-fitting method. The effect of increasing chloride ion concentration on melting profiles was 2-fold. Below 0.6 M salt, the two transition temperatures, Tm,1 and Tm,2, depended on salt concentration such that increasing NaCl concentration caused a small stabilization of the helix while increasing acetate concentration caused the helix to become markedly more stable. Between 0.6 and 1.0 M, variation of chloride concentration had almost no effect on the thermal stability of the rod while increasing acetate concentration increased its stability considerably. Above 1.0 M NaCl, the melting profiles became broad with a third transition being observed (e.g., at 3.0 M, Tm,3 = 38 degrees C), indicating the existence of a region which has a tendency to be destabilized by chloride. The third transition was not observed at comparable concentrations of acetate. This effect of chloride was not expected on the basis of its position in the Hofmeister series.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D000838:Anions; D002942:Circular Dichroism; D004355:Drug Stability; D007700:Kinetics; D008433:Mathematics; D008954:Models, Biological; D009132:Muscles / Q000378:metabolism*; D009218:Myosins / Q000378:metabolism*; D010206:Papain / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis; D011487:Protein Conformation; D011817:Rabbits; D013816:Thermodynamics
D000838:Anions; D010446:Peptide Fragments; D010206:Papain; D009218:Myosins