Conventional and saturation transfer EPR spectroscopy of Na+/K+-ATPase modified with different maleimide-nitroxide derivatives

Mikael Esmann, Kálmán Hideg, Derek Marsh

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The membranous Na+/K+-ATPase from Squalus acanthias has been covalently modified on either Class I or Class II sulphydryl groups using derivatives of 3-(maleimidomethyl)-1-oxyl-2,2,5,5,-tetramethylpyrrolidine with substituents of different charge and hydrophobicity attached at the remaining unsubstituted position of the pyrrolidine ring. The substituent groups used were a methyl and a hexyl ester, and di- and tri-methylammonium ethyl esters, as well as the parent underivatized compound. Additionally, another series of maleimide-nitroxides differing (by zero to seven intervening atoms) in the length of the linking group between the maleimide and the pyrrolidine moieties was used. The sites of attachment have been characterized in terms of the rotational mobility and environment polarity by using conventional and saturation transfer EPR spectroscopy of these spin-labelled reagents. This provides a further sub-classification of the primary Class I and Class II SH-groups on the α-subunit of the enzyme, which differ both in their reactivity and influence on the Na+/K+-ATPase activity.

Original languageEnglish
Pages (from-to)51-59
Number of pages9
JournalBiochimica et Biophysica Acta (BBA)/Protein Structure and Molecular
Volume1159
Issue number1
DOIs
Publication statusPublished - Sep 4 1992

Fingerprint

Paramagnetic resonance
Adenosine Triphosphatases
Spectrum Analysis
Esters
Squalus acanthias
Spectroscopy
Derivatives
Hydrophobicity
Hydrophobic and Hydrophilic Interactions
Atoms
Enzymes
nitroxide maleimide
pyrrolidine
methylamine
sodium-translocating ATPase
2,2,5,5,-tetramethylpyrrolidine-1-oxyl
maleimide

Keywords

  • (S. acanthias)
  • ATPase
  • EPR
  • Na/K

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Structural Biology

Cite this

@article{dcee9454106a4c2d8f568bb4cffecd88,
title = "Conventional and saturation transfer EPR spectroscopy of Na+/K+-ATPase modified with different maleimide-nitroxide derivatives",
abstract = "The membranous Na+/K+-ATPase from Squalus acanthias has been covalently modified on either Class I or Class II sulphydryl groups using derivatives of 3-(maleimidomethyl)-1-oxyl-2,2,5,5,-tetramethylpyrrolidine with substituents of different charge and hydrophobicity attached at the remaining unsubstituted position of the pyrrolidine ring. The substituent groups used were a methyl and a hexyl ester, and di- and tri-methylammonium ethyl esters, as well as the parent underivatized compound. Additionally, another series of maleimide-nitroxides differing (by zero to seven intervening atoms) in the length of the linking group between the maleimide and the pyrrolidine moieties was used. The sites of attachment have been characterized in terms of the rotational mobility and environment polarity by using conventional and saturation transfer EPR spectroscopy of these spin-labelled reagents. This provides a further sub-classification of the primary Class I and Class II SH-groups on the α-subunit of the enzyme, which differ both in their reactivity and influence on the Na+/K+-ATPase activity.",
keywords = "(S. acanthias), ATPase, EPR, Na/K",
author = "Mikael Esmann and K{\'a}lm{\'a}n Hideg and Derek Marsh",
year = "1992",
month = "9",
day = "4",
doi = "10.1016/0167-4838(92)90074-N",
language = "English",
volume = "1159",
pages = "51--59",
journal = "Biochimica et Biophysica Acta - Proteins and Proteomics",
issn = "1570-9639",
publisher = "Elsevier",
number = "1",

}

TY - JOUR

T1 - Conventional and saturation transfer EPR spectroscopy of Na+/K+-ATPase modified with different maleimide-nitroxide derivatives

AU - Esmann, Mikael

AU - Hideg, Kálmán

AU - Marsh, Derek

PY - 1992/9/4

Y1 - 1992/9/4

N2 - The membranous Na+/K+-ATPase from Squalus acanthias has been covalently modified on either Class I or Class II sulphydryl groups using derivatives of 3-(maleimidomethyl)-1-oxyl-2,2,5,5,-tetramethylpyrrolidine with substituents of different charge and hydrophobicity attached at the remaining unsubstituted position of the pyrrolidine ring. The substituent groups used were a methyl and a hexyl ester, and di- and tri-methylammonium ethyl esters, as well as the parent underivatized compound. Additionally, another series of maleimide-nitroxides differing (by zero to seven intervening atoms) in the length of the linking group between the maleimide and the pyrrolidine moieties was used. The sites of attachment have been characterized in terms of the rotational mobility and environment polarity by using conventional and saturation transfer EPR spectroscopy of these spin-labelled reagents. This provides a further sub-classification of the primary Class I and Class II SH-groups on the α-subunit of the enzyme, which differ both in their reactivity and influence on the Na+/K+-ATPase activity.

AB - The membranous Na+/K+-ATPase from Squalus acanthias has been covalently modified on either Class I or Class II sulphydryl groups using derivatives of 3-(maleimidomethyl)-1-oxyl-2,2,5,5,-tetramethylpyrrolidine with substituents of different charge and hydrophobicity attached at the remaining unsubstituted position of the pyrrolidine ring. The substituent groups used were a methyl and a hexyl ester, and di- and tri-methylammonium ethyl esters, as well as the parent underivatized compound. Additionally, another series of maleimide-nitroxides differing (by zero to seven intervening atoms) in the length of the linking group between the maleimide and the pyrrolidine moieties was used. The sites of attachment have been characterized in terms of the rotational mobility and environment polarity by using conventional and saturation transfer EPR spectroscopy of these spin-labelled reagents. This provides a further sub-classification of the primary Class I and Class II SH-groups on the α-subunit of the enzyme, which differ both in their reactivity and influence on the Na+/K+-ATPase activity.

KW - (S. acanthias)

KW - ATPase

KW - EPR

KW - Na/K

UR - http://www.scopus.com/inward/record.url?scp=0026698710&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026698710&partnerID=8YFLogxK

U2 - 10.1016/0167-4838(92)90074-N

DO - 10.1016/0167-4838(92)90074-N

M3 - Article

C2 - 1327155

AN - SCOPUS:0026698710

VL - 1159

SP - 51

EP - 59

JO - Biochimica et Biophysica Acta - Proteins and Proteomics

JF - Biochimica et Biophysica Acta - Proteins and Proteomics

SN - 1570-9639

IS - 1

ER -