title: [Effect of ligands on rotational mobility of Na,K-ATPase].
abstract: Phosphorescence anisotropy of eosin-5'-isothiocyanate labelled Na,K-ATPase purified from duck salt glands has been studied. The initial anisotropy value is 0.235 +/- 0.015 (room temperature) and does not depend on the enzyme conformation (sodium or potassium). The experimental curve is fitted into a two-exponential curve with residual term, the fast component corresponds to the rotational mobility of the functional unit of Na,K-ATPase (promoter), while the slow one--to that of larger associates. In the presence of ligands modifying the conformational state of Na,K-ATPase (sodium, potassium, ATP) the rotational mobility of the fast component does not change in contrast with the slow one. A comparison of the enzyme rotational mobility in the presence of ligands simulating different steps of hydrolytic cycle suggests that interprotomer interactions are changed in the course of hydrolytic cycle: the fraction of larger associates increases at the step of the enzyme interactions with potassium ions, whereas their mobility in the bilayer enhances sharply after interaction with ATP. In the presence of the 2% non-ionic detergent, C12E9, the initial anisotropy value decreases down to 0.1; the residual term disappears thereby, while the curve is still two-exponential. However, the difference in the rotational mobility of sodium and potassium conformers diminishes. At the same time, the ratios between protomers and oligomers in the presence of sodium and potassium become approximated. This indicates that in the presence of the detergent high molecular weight associates are solubilized, the mobility of the both protomers and oligomers of Na,K-ATPase increases, while the difference between the mobilities of sodium and potassium conformers is disappeared.
authors.affiliation: School of Applied Sciences, Robert Gordon University, Aberdeen, Scotland, UK.
authors.forename: A A
authors.initials: AA
authors.lastname: Boldyrev
authors.forename: A M
authors.initials: AM
authors.lastname: Rubtsov
authors.forename: O D
authors.initials: OD
authors.lastname: Lopina
authors.forename: D
authors.initials: D
authors.lastname: McStay
authors.forename: L
authors.initials: L
authors.lastname: Yang
authors.forename: P J
authors.initials: PJ
authors.lastname: Quinn
mesh_terms: D000818:Animals; D001704:Biopolymers; D004372:Ducks; D006868:Hydrolysis; D008024:Ligands; D011487:Protein Conformation; D012491:Salt Gland / Q000201:enzymology; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000378:metabolism*
chemical_list: D001704:Biopolymers; D008024:Ligands; D000254:Sodium-Potassium-Exchanging ATPase
vernacular_title: Vliianie ligandov na vrashchatel'nuiu podvizhnost' Na,K-ATPazy.