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Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements

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Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements. / Schwappach, B; Stürmer, W; Apell, H J; Karlish, S J.

in: J BIOL CHEM, Jahrgang 269, Nr. 34, 26.08.1994, S. 21620-6.

Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung

Harvard

Schwappach, B, Stürmer, W, Apell, HJ & Karlish, SJ 1994, 'Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements', J BIOL CHEM, Jg. 269, Nr. 34, S. 21620-6.

APA

Schwappach, B., Stürmer, W., Apell, H. J., & Karlish, S. J. (1994). Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements. J BIOL CHEM, 269(34), 21620-6.

Vancouver

Schwappach B, Stürmer W, Apell HJ, Karlish SJ. Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements. J BIOL CHEM. 1994 Aug 26;269(34):21620-6.

Bibtex

@article{b66dfe37981d400e98187af2aa9e095d,
title = "Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements",
abstract = "A fluorescent dye, RH421, has been used to characterize charge movements associated with cation and cardiotonic steroid binding to Na,K-ATPase and to a specifically trypsinized preparation, so-called {"}19-kDa membranes.{"} A fluorescence decrease induced by Na+ is attributed to electrogenic binding of one Na+ ion from the cytoplasm. The apparent affinity for Na+ is the same in both preparations. (ATP + Na + Mg) or (P(i) + Mg)-induced fluorescence signals observed with native enzyme are not observed in 19-kDa membranes, consistent with loss of ATP binding and phosphorylation. Cardiotonic steroids (CS) bind to native enzyme and 19-kDa membranes as judged by RH421 signals, fluorescence of anthroyl ouabain, and inhibition of Rb+ occlusion. Binding affinities to both preparations are in the micromolar range, and binding is prevented by the presence of Na+ or K+. The kinetics of glycone binding and dissociation are identical in both preparations, but aglycones bind and dissociate about 6-fold faster to 19-kDa membranes. Binding of Na+ and cardiotonic steroids is inactivated upon heating or extensive Pronase digestion of 19-kDa membranes. This suggests that cation and CS binding depend on the structural integrity of a complex of the proteolytic fragments, and that sites for both cations or CS consist of ligating groups located on more than one fragments of 19-kDa membranes.",
keywords = "Animals, Cardiac Glycosides/metabolism, Cations, Monovalent/metabolism, Cell Membrane/metabolism, Electricity, Fluorescent Dyes/metabolism, Kidney Medulla/metabolism, Models, Biological, Motion, Peptide Fragments/metabolism, Pyridinium Compounds/metabolism, Rabbits, Rubidium/metabolism, Sodium/metabolism, Sodium-Potassium-Exchanging ATPase/drug effects, Spectrometry, Fluorescence, Styrenes/metabolism, Trypsin/pharmacology",
author = "B Schwappach and W St{\"u}rmer and Apell, {H J} and Karlish, {S J}",
year = "1994",
month = aug,
day = "26",
language = "English",
volume = "269",
pages = "21620--6",
journal = "J BIOL CHEM",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "34",

}

RIS

TY - JOUR

T1 - Binding of sodium ions and cardiotonic steroids to native and selectively trypsinized Na,K pump, detected by charge movements

AU - Schwappach, B

AU - Stürmer, W

AU - Apell, H J

AU - Karlish, S J

PY - 1994/8/26

Y1 - 1994/8/26

N2 - A fluorescent dye, RH421, has been used to characterize charge movements associated with cation and cardiotonic steroid binding to Na,K-ATPase and to a specifically trypsinized preparation, so-called "19-kDa membranes." A fluorescence decrease induced by Na+ is attributed to electrogenic binding of one Na+ ion from the cytoplasm. The apparent affinity for Na+ is the same in both preparations. (ATP + Na + Mg) or (P(i) + Mg)-induced fluorescence signals observed with native enzyme are not observed in 19-kDa membranes, consistent with loss of ATP binding and phosphorylation. Cardiotonic steroids (CS) bind to native enzyme and 19-kDa membranes as judged by RH421 signals, fluorescence of anthroyl ouabain, and inhibition of Rb+ occlusion. Binding affinities to both preparations are in the micromolar range, and binding is prevented by the presence of Na+ or K+. The kinetics of glycone binding and dissociation are identical in both preparations, but aglycones bind and dissociate about 6-fold faster to 19-kDa membranes. Binding of Na+ and cardiotonic steroids is inactivated upon heating or extensive Pronase digestion of 19-kDa membranes. This suggests that cation and CS binding depend on the structural integrity of a complex of the proteolytic fragments, and that sites for both cations or CS consist of ligating groups located on more than one fragments of 19-kDa membranes.

AB - A fluorescent dye, RH421, has been used to characterize charge movements associated with cation and cardiotonic steroid binding to Na,K-ATPase and to a specifically trypsinized preparation, so-called "19-kDa membranes." A fluorescence decrease induced by Na+ is attributed to electrogenic binding of one Na+ ion from the cytoplasm. The apparent affinity for Na+ is the same in both preparations. (ATP + Na + Mg) or (P(i) + Mg)-induced fluorescence signals observed with native enzyme are not observed in 19-kDa membranes, consistent with loss of ATP binding and phosphorylation. Cardiotonic steroids (CS) bind to native enzyme and 19-kDa membranes as judged by RH421 signals, fluorescence of anthroyl ouabain, and inhibition of Rb+ occlusion. Binding affinities to both preparations are in the micromolar range, and binding is prevented by the presence of Na+ or K+. The kinetics of glycone binding and dissociation are identical in both preparations, but aglycones bind and dissociate about 6-fold faster to 19-kDa membranes. Binding of Na+ and cardiotonic steroids is inactivated upon heating or extensive Pronase digestion of 19-kDa membranes. This suggests that cation and CS binding depend on the structural integrity of a complex of the proteolytic fragments, and that sites for both cations or CS consist of ligating groups located on more than one fragments of 19-kDa membranes.

KW - Animals

KW - Cardiac Glycosides/metabolism

KW - Cations, Monovalent/metabolism

KW - Cell Membrane/metabolism

KW - Electricity

KW - Fluorescent Dyes/metabolism

KW - Kidney Medulla/metabolism

KW - Models, Biological

KW - Motion

KW - Peptide Fragments/metabolism

KW - Pyridinium Compounds/metabolism

KW - Rabbits

KW - Rubidium/metabolism

KW - Sodium/metabolism

KW - Sodium-Potassium-Exchanging ATPase/drug effects

KW - Spectrometry, Fluorescence

KW - Styrenes/metabolism

KW - Trypsin/pharmacology

M3 - SCORING: Journal article

C2 - 8063803

VL - 269

SP - 21620

EP - 21626

JO - J BIOL CHEM

JF - J BIOL CHEM

SN - 0021-9258

IS - 34

ER -