A particle-bound, Mg2+ -dependent ATPase activity is investigated in cell-free extracts of the tyanobacterium Anabaena variabilis. The enzyme can be clearly distinguished from the cyanobacterial coupling factor of photophosphorylation and from the alkaline phosphatase. It requires low concentrations of Ca2+ for maximal activity and is inhibited by ortho-vanadate, indicating that the enzyme may form a phosphorylated intermediate in its catalytic cycle. The distribution of the ATPase in sucrose density gradients does not follow that of thylakoids. It is concluded from these characteristics that the enzyme is bound to the plasma membrane. The cytochrome oxidase activity of the extracts appears to be restricted to the thylakoids.