Characterization of a proton pump from Acer pseudoplatanus cell microsomes

Abstract An Acer pseudoplatanus cell microsomal fraction was enriched in ATPase by sedimentation through a sucrose cushion and treatment with Triton X-100. This activity, which reached 0.9 μmol P i min −1 mg −1 protein, was specific for ATP, slightly stimulated by K + , inhibited by orthovanadate and diethylstilbestrol, insensitive to oligomycin and azide, and had a K m - value of 0.51 mM for MgATP. ATP-dependent proton translocation was demonstrated by the ΔpH probe acridine orange. This activity had a optimum at pH 6.5, was substrate specific for ATP, and was strongly dependent on K + . Preparations of plasma membrane ATPase from A. pseudoplatanus cell culture thus posses biochemical properties analogous to other well characterized plant plasma membrane enzymes.