Activation of dynein 1 adenosine triphosphatase by monovalent salts and inhibition by vanadate.

The ATPase activity of native dynein 1 from sea urchin sperm flagella is activated reversibly by inorganic monovalent chlorides with the magnitude of activation being nearly independent of the cation below 0.3 M. At higher concentrations, activation increases in the order LiCl greater than NH4Cl greater than NaCl greater than KCl, with the maximum occurring at about 0.8 M in all cases. The sodium halides activate reversibly in the order NaI greater than NaBr greater than NaCl, but NaF is strongly inhibitory. The presence of the organic anions formate, acetate, or propionate favors the native low ATPase activity state, with lithium acetate giving little activation at up to 1 M and sodium acetate partially reversing the activation due to simultaneous presence of 0.6 M NaCl. The sedimentation rate of the dynein does not change between 0.2 and 0.8 M NaCl or sodium acetate, suggesting that the effects of the anions on ATPase activity are due to local changes near the catalytic site, rather than to large-scale changes in the molecular structure. All the agents that activate the dynein ATPase, either reversibly (halides) or irreversibly (Triton X-100), decrease its sensitivity to inhibition by vanadate, consistent with ATPase activation being the result of a decreased stability of the dynein. ADP.Pi kinetic intermediate that is thought to bind vanadate at the gamma-Pi site and act as a dead-end kinetic block. Although many divalent cations, including Mg2+, Mn2+, Fe2+, Co2+, Ni2+, Zn2+, Ca2+, and Sr2+, can support dynein ATPase activity, the magnitude of ATPase increase observed upon treatment with Triton X-100 is greatest with Mg2+ and Mn2+, which are also the only cations capable of supporting the motility of demembranated flagella at rates similar to those observed in vivo.