Structural identification of cation binding pockets in the plasma membrane proton pump.

Abstract

The activity of P-type plasma membrane H(+)-ATPases is modulated by H(+) and cations, with K(+) and Ca(2+) being of physiological relevance. Using X-ray crystallography, we have located the binding site for Rb(+) as a K(+) congener, and for Tb(3+) and Ho(3+) as Ca(2+) congeners. Rb(+) is found coordinated by a conserved aspartate residue in the phosphorylation domain. A single Tb(3+) ion is identified positioned in the nucleotide-binding domain in close vicinity to the bound nucleotide. Ho(3+) ions are coordinated at two distinct sites within the H(+)-ATPase: One site is at the interface of the nucleotide-binding and phosphorylation domains, and the other is in the transmembrane domain toward the extracellular side. The identified binding sites are suggested to represent binding pockets for regulatory cations and a H(+) binding site for protons leaving the pump molecule. This implicates Ho(3+) as a novel chemical tool for identification of proton binding sites.