title: Molecular mechanism of Na(+),K(+)-ATPase malfunction in mutations characteristic of adrenal hypertension.
abstract: Mutations within ion-transporting proteins may severely affect their ability to traffic ions properly and thus perturb the delicate balance of ion gradients. Somatic gain-of-function mutations of the Na(+),K(+)-ATPase α1-subunit have been found in aldosterone-producing adenomas that are among the causes of hypertension. We used molecular dynamics simulations to investigate the structural consequences of these mutations, namely, Leu97 substitution by Arg (L97R), Val325 substitution by Gly (V325G), deletion of residues 93-97 (Del93-97), and deletion-substitution of residues 953-956 by Ser (EETA956S), which shows inward leak currents under physiological conditions. The first three mutations affect the structural context of the key ion-binding residue Glu327 at binding site II, which leads to the loss of the ability to bind ions correctly and to occlude the pump. The mutated residue in L97R is more hydrated, which ultimately leads to the observed proton leak. V325G mimics the structural behavior of L97R; however, it does not promote the hydration of surrounding residues. In Del93-97, a broader opening is observed because of the rearrangement of the kinked transmembrane helix 1, M1, which may explain the sodium leak measured with the mutant. The last mutant, EETA956S, opens an additional water pathway near the C-terminus, affecting the III sodium-specific binding site. The results are in excellent agreement with recent electrophysiology measurements and suggest how three mutations prevent the occlusion of the Na(+),K(+)-ATPase, with the possibility of transforming the pump into a passive ion channel, whereas the fourth mutation provides insight into the sodium binding in the E1 state.
authors.affiliation: MEMPHYS-Center for Biomembrane Physics, University of Southern Denmark , Campusvej 55, DK-5230 Odense M, Denmark.
authors.forename: Wojciech
authors.initials: W
authors.lastname: Kopec
authors.forename: Bastien
authors.initials: B
authors.lastname: Loubet
authors.forename: Hanne
authors.initials: H
authors.lastname: Poulsen
authors.forename: Himanshu
authors.initials: H
authors.lastname: Khandelia
mesh_terms: D000307:Adrenal Gland Diseases / Q000150:complications / Q000235:genetics*; D001665:Binding Sites; D002414:Cations, Monovalent; D006801:Humans; D006973:Hypertension / Q000209:etiology / Q000235:genetics*; D056004:Molecular Dynamics Simulation; D009154:Mutation; D010042:Ouabain / Q000737:chemistry; D011485:Protein Binding; D012964:Sodium / Q000737:chemistry; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry* / Q000235:genetics; D055672:Static Electricity; D014867:Water / Q000737:chemistry
chemical_list: D002414:Cations, Monovalent; D014867:Water; D010042:Ouabain; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase