Rotational mobility of Ca+-ATPase of sarcoplasmic reticulum in viscous media

Marianna Török, Györgyi Jakab, Alajos Bérczi, László Dux, László I. Horváth

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

The rotational diffusion of Ca2+-ATPase [Ca2+,Mg2+-activated ATP phosphohydrolase E.C. 3.6.1.38] was studied in native sarcoplasmic reticulum membrane by saturation transfer ESR spectroscopy after covalent labelling of intramembranous sulfhydryl groups with nitroxyl derivative of maleimide (5-MSL) as a function of sucrose and glycerol in the suspending medium. The relative enzymatic activity of sarcoplasmic reticulum was followed by increasing the viscosity of the aqueous phase. The ATP hydrolysing activity of the enzyme decreased differently on adding sucrose and glycerol. In the case of sucrose the reciprocal of power dependence of viscosity was observed, whereas for glycerol an exponential decay law was obtained, indicating solvent-protein interaction. On increasing the viscosity of the aqueous phase by either sucrose or glycerol, no changes were observed in the intramembranous viscosity as measured using intercalated spin-labelled stearic acid (16-SASL). The effective rotational correlation time of the protein was measured, as a mobility parameter,using saturation transfer ESR spectroscopy and found to be increased linearly with the viscosity of the sucrose containing medium and for the extramembranous size a height of 6.8 nm was obtained, indicating that approx. 82% of the volume of Ca2+-ATPase protein is external to the sarcoplasmic reticulum. The addition of glycerol probably promoted protein-protein interaction, as indicated by the larger changes in rotational diffusion and non-linear viscosity dependence.

Original languageEnglish
Pages (from-to)193-200
Number of pages8
JournalBBA - Biomembranes
Volume1326
Issue number2
DOIs
Publication statusPublished - Jun 12 1997

Fingerprint

Sarcoplasmic Reticulum
Viscosity
Adenosine Triphosphatases
Glycerol
Sucrose
Proteins
Paramagnetic resonance
Spectrum Analysis
Spectroscopy
Labeling
Adenosine Triphosphate
Derivatives
Membranes
Enzymes

Keywords

  • Ca-ATPase
  • Rotational mobility
  • Saturation transfer ESR
  • Viscosity dependence

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology

Cite this

Rotational mobility of Ca+-ATPase of sarcoplasmic reticulum in viscous media. / Török, Marianna; Jakab, Györgyi; Bérczi, Alajos; Dux, László; Horváth, László I.

In: BBA - Biomembranes, Vol. 1326, No. 2, 12.06.1997, p. 193-200.

Research output: Contribution to journalArticle

Török, Marianna ; Jakab, Györgyi ; Bérczi, Alajos ; Dux, László ; Horváth, László I. / Rotational mobility of Ca+-ATPase of sarcoplasmic reticulum in viscous media. In: BBA - Biomembranes. 1997 ; Vol. 1326, No. 2. pp. 193-200.
@article{7729a7026bc64e36922d07faa1ca69ad,
title = "Rotational mobility of Ca+-ATPase of sarcoplasmic reticulum in viscous media",
abstract = "The rotational diffusion of Ca2+-ATPase [Ca2+,Mg2+-activated ATP phosphohydrolase E.C. 3.6.1.38] was studied in native sarcoplasmic reticulum membrane by saturation transfer ESR spectroscopy after covalent labelling of intramembranous sulfhydryl groups with nitroxyl derivative of maleimide (5-MSL) as a function of sucrose and glycerol in the suspending medium. The relative enzymatic activity of sarcoplasmic reticulum was followed by increasing the viscosity of the aqueous phase. The ATP hydrolysing activity of the enzyme decreased differently on adding sucrose and glycerol. In the case of sucrose the reciprocal of power dependence of viscosity was observed, whereas for glycerol an exponential decay law was obtained, indicating solvent-protein interaction. On increasing the viscosity of the aqueous phase by either sucrose or glycerol, no changes were observed in the intramembranous viscosity as measured using intercalated spin-labelled stearic acid (16-SASL). The effective rotational correlation time of the protein was measured, as a mobility parameter,using saturation transfer ESR spectroscopy and found to be increased linearly with the viscosity of the sucrose containing medium and for the extramembranous size a height of 6.8 nm was obtained, indicating that approx. 82{\%} of the volume of Ca2+-ATPase protein is external to the sarcoplasmic reticulum. The addition of glycerol probably promoted protein-protein interaction, as indicated by the larger changes in rotational diffusion and non-linear viscosity dependence.",
keywords = "Ca-ATPase, Rotational mobility, Saturation transfer ESR, Viscosity dependence",
author = "Marianna T{\"o}r{\"o}k and Gy{\"o}rgyi Jakab and Alajos B{\'e}rczi and L{\'a}szl{\'o} Dux and Horv{\'a}th, {L{\'a}szl{\'o} I.}",
year = "1997",
month = "6",
day = "12",
doi = "10.1016/S0005-2736(97)00021-7",
language = "English",
volume = "1326",
pages = "193--200",
journal = "Biochimica et Biophysica Acta - Biomembranes",
issn = "0005-2736",
publisher = "Elsevier",
number = "2",

}

TY - JOUR

T1 - Rotational mobility of Ca+-ATPase of sarcoplasmic reticulum in viscous media

AU - Török, Marianna

AU - Jakab, Györgyi

AU - Bérczi, Alajos

AU - Dux, László

AU - Horváth, László I.

PY - 1997/6/12

Y1 - 1997/6/12

N2 - The rotational diffusion of Ca2+-ATPase [Ca2+,Mg2+-activated ATP phosphohydrolase E.C. 3.6.1.38] was studied in native sarcoplasmic reticulum membrane by saturation transfer ESR spectroscopy after covalent labelling of intramembranous sulfhydryl groups with nitroxyl derivative of maleimide (5-MSL) as a function of sucrose and glycerol in the suspending medium. The relative enzymatic activity of sarcoplasmic reticulum was followed by increasing the viscosity of the aqueous phase. The ATP hydrolysing activity of the enzyme decreased differently on adding sucrose and glycerol. In the case of sucrose the reciprocal of power dependence of viscosity was observed, whereas for glycerol an exponential decay law was obtained, indicating solvent-protein interaction. On increasing the viscosity of the aqueous phase by either sucrose or glycerol, no changes were observed in the intramembranous viscosity as measured using intercalated spin-labelled stearic acid (16-SASL). The effective rotational correlation time of the protein was measured, as a mobility parameter,using saturation transfer ESR spectroscopy and found to be increased linearly with the viscosity of the sucrose containing medium and for the extramembranous size a height of 6.8 nm was obtained, indicating that approx. 82% of the volume of Ca2+-ATPase protein is external to the sarcoplasmic reticulum. The addition of glycerol probably promoted protein-protein interaction, as indicated by the larger changes in rotational diffusion and non-linear viscosity dependence.

AB - The rotational diffusion of Ca2+-ATPase [Ca2+,Mg2+-activated ATP phosphohydrolase E.C. 3.6.1.38] was studied in native sarcoplasmic reticulum membrane by saturation transfer ESR spectroscopy after covalent labelling of intramembranous sulfhydryl groups with nitroxyl derivative of maleimide (5-MSL) as a function of sucrose and glycerol in the suspending medium. The relative enzymatic activity of sarcoplasmic reticulum was followed by increasing the viscosity of the aqueous phase. The ATP hydrolysing activity of the enzyme decreased differently on adding sucrose and glycerol. In the case of sucrose the reciprocal of power dependence of viscosity was observed, whereas for glycerol an exponential decay law was obtained, indicating solvent-protein interaction. On increasing the viscosity of the aqueous phase by either sucrose or glycerol, no changes were observed in the intramembranous viscosity as measured using intercalated spin-labelled stearic acid (16-SASL). The effective rotational correlation time of the protein was measured, as a mobility parameter,using saturation transfer ESR spectroscopy and found to be increased linearly with the viscosity of the sucrose containing medium and for the extramembranous size a height of 6.8 nm was obtained, indicating that approx. 82% of the volume of Ca2+-ATPase protein is external to the sarcoplasmic reticulum. The addition of glycerol probably promoted protein-protein interaction, as indicated by the larger changes in rotational diffusion and non-linear viscosity dependence.

KW - Ca-ATPase

KW - Rotational mobility

KW - Saturation transfer ESR

KW - Viscosity dependence

UR - http://www.scopus.com/inward/record.url?scp=0343990106&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0343990106&partnerID=8YFLogxK

U2 - 10.1016/S0005-2736(97)00021-7

DO - 10.1016/S0005-2736(97)00021-7

M3 - Article

VL - 1326

SP - 193

EP - 200

JO - Biochimica et Biophysica Acta - Biomembranes

JF - Biochimica et Biophysica Acta - Biomembranes

SN - 0005-2736

IS - 2

ER -