title: [Novel ratchet mechanism of gastric H(+), K(+)-ATPase revealed by electron crystallography of two-dimensional crystals].
abstract: Acid secretion by the stomach results in a pH of about 1. This highly acidic environment is essential for digestion and also acts as a first barrier against bacterial and viral infections. Conversely, too much acid secretion causes gastric ulcer. The mechanism by which this massive proton gradient is generated is of considerable biomedical interest. In this review, we introduce the first molecular model for this remarkable biological phenomenon. The structure of H(+),K(+)-ATPase at 6.5 A resolution was determined by electron crystallography of two-dimensional crystals. The structure shows the catalytic alpha-subunit and the non-catalytic beta-subunit in a pseudo-E(2)P conformation. Different from Na(+),K(+)-ATPase, the N-terminal tail of the beta-subunit is in direct contact with the phosphorylation domain of the alpha-subunit. This interaction may hold the phosphorylation domain in place, thus stabilizing the enzyme conformation and preventing the reverse reaction of the transport cycle. Indeed, truncation of the beta-subunit N-terminus allowed the reverse reaction to occur. These results suggest that the N-terminal tail of the beta-subunit functions as a "ratchet", preventing inefficient transport and reverse-flow of protons. We can thus provide a mechanistic explanation for how the H(+),K(+)-ATPase can generate a million-fold proton gradient across the gastric parietal cell membrane, the highest cation gradient known in any mammalian tissue.
authors.affiliation: Department of Biophysics, Faculty of Science, Kyoto University, Kyoto, Japan. ikkei@em.biophys.kyoto-u.ac.jp
authors.forename: Kazuhiro
authors.initials: K
authors.lastname: Abe
authors.forename: Kazutoshi
authors.initials: K
authors.lastname: Tani
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Nishizawa
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Fujiyoshi
mesh_terms: D000818:Animals; D002384:Catalysis; D003460:Crystallization; D003461:Crystallography; D004583:Electrons; D005753:Gastric Mucosa / Q000201:enzymology / Q000378:metabolism; D017506:H(+)-K(+)-Exchanging ATPase / Q000737:chemistry* / Q000502:physiology*; D006801:Humans; D010766:Phosphorylation; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011522:Protons
chemical_list: D011522:Protons; D017506:H(+)-K(+)-Exchanging ATPase