title: Na,K-ATPase labelled with 5-iodoacetamidofluorescein: E2-E1 conformational transition induced by different nucleotides.
abstract: A conformational transition between E2 and E1 forms of Na, K-ATPase induced by different nucleotides has been studied under steady state conditions using the enzyme labelled with 5-iodoacetamidofluorescein. In the presence of K+ the plot of fluorescence as a function of [ATP], [ADP] or [CTP] (in a range of 5 microM-12 mM) is a biphasic one. A similar dependence for AMP, ITP, GTP and UTP demonstrates a hyperbolic behaviour. The data suggest that the shift in the equilibrium between E2 and E1 forms of Na,K-ATPase towards the E1 conformation is induced by ATP binding both with high and low affinity sites. Two structural features of ATP are apparently important for its interaction with more than one type of ATP binding sites or for providing for E2-E1 transition induced by this interaction: (i) beta-phosphate group in the terminal part of the molecule, (ii) unprotonated N1 and/or NH2-group in the 6th position of the purine base.
authors.affiliation: Department of Biochemistry, School of Biology, Moscow State University, USSR.
authors.forename: O D
authors.initials: OD
authors.lastname: Lopina
authors.forename: E V
authors.initials: EV
authors.lastname: Pindel
authors.forename: A A
authors.initials: AA
authors.lastname: Boldyrev
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001665:Binding Sites; D004372:Ducks; D005452:Fluoresceins; D008274:Magnesium / Q000494:pharmacology; D009711:Nucleotides / Q000378:metabolism*; D010042:Ouabain / Q000494:pharmacology; D011189:Potassium Chloride; D011487:Protein Conformation; D012965:Sodium Chloride; D000254:Sodium-Potassium-Exchanging ATPase / Q000378:metabolism; D013050:Spectrometry, Fluorescence
chemical_list: D005452:Fluoresceins; D009711:Nucleotides; D012965:Sodium Chloride; D010042:Ouabain; C015296:5-iodoacetamidofluorescein; D011189:Potassium Chloride; D000255:Adenosine Triphosphate; D000254:Sodium-Potassium-Exchanging ATPase; D008274:Magnesium