Role of sulfhydryl groups in phospholipid methylation reactions of cardiac sarcolemma

Roland Vetter, Jian Dai, Nasrin Mesaeli, Vincenzo Panagia, Naranjan S. Dhalla

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

The effect of reagents that modify sulfur-containing amino acid residues in the phosphatidylethanolamine N-methyltransferase was studied in the isolated rat cardiac sarcolemma by employing S-adenosyl-L-[methyl-3H]methionine as a methyl donor. Dithiothreitol protected the sulfhydryl groups in the membrane and caused a concentration- and time-dependent increase of phospholipid N-methylation at three different catalytic sites. This stimulation was highest (9-fold) in the presence of 1 MM MgCl2 and 0.1 μM S-adenosyl-L-[methyl-3H]methionine at pH 8.0 (catalytic site 1), and was associated with an enhancement of Vmax without changes in Km for the methyl donor. Thiol glutathione was less stimulatory than dithiothreitol; glutathione disulfide inhibited the phosphatidylethanolamine N-methylation by 50%. The alkylating reagents, N-ethylmaleimide and methylmethanethiosulfonate, inhibited the N-methylation with IC5O of 6.9 and 14.1 μM, respectively; this inhibition was prevented by 1 mM dithiothreitol. These results indicate a critical role of sulfhydryl groups for the activity of the cardiac sarcolemmal phosphatidylethanolamine N-methyltransferase and suggest that this enzyme system in cardiac sarcolemma may be controlled by the glutathione/glutathione disulfide redox state in the cell.

Original languageEnglish
Pages (from-to)85-96
Number of pages12
JournalMolecular and Cellular Biochemistry
Volume103
Issue number1
DOIs
Publication statusPublished - Apr 1991
Externally publishedYes

Fingerprint

Sarcolemma
Methylation
Dithiothreitol
Phosphatidylethanolamine N-Methyltransferase
Phospholipids
Glutathione Disulfide
Glutathione
Catalytic Domain
Sulfur Amino Acids
Magnesium Chloride
Ethylmaleimide
Sulfur
Sulfhydryl Compounds
Oxidation-Reduction
Rats
Membranes
Amino Acids
Enzymes
methionine methyl ester

Keywords

  • neutral lipid methylation
  • phospholipid N-methylation
  • rat heart
  • S-adenosyl-L-methionine
  • sarcolemma
  • sulfhydryl groups

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Clinical Biochemistry
  • Cell Biology

Cite this

Role of sulfhydryl groups in phospholipid methylation reactions of cardiac sarcolemma. / Vetter, Roland; Dai, Jian; Mesaeli, Nasrin; Panagia, Vincenzo; Dhalla, Naranjan S.

In: Molecular and Cellular Biochemistry, Vol. 103, No. 1, 04.1991, p. 85-96.

Research output: Contribution to journalArticle

Vetter, Roland ; Dai, Jian ; Mesaeli, Nasrin ; Panagia, Vincenzo ; Dhalla, Naranjan S. / Role of sulfhydryl groups in phospholipid methylation reactions of cardiac sarcolemma. In: Molecular and Cellular Biochemistry. 1991 ; Vol. 103, No. 1. pp. 85-96.
@article{1ecf9ea668c742e3b28de05e6d6a08f3,
title = "Role of sulfhydryl groups in phospholipid methylation reactions of cardiac sarcolemma",
abstract = "The effect of reagents that modify sulfur-containing amino acid residues in the phosphatidylethanolamine N-methyltransferase was studied in the isolated rat cardiac sarcolemma by employing S-adenosyl-L-[methyl-3H]methionine as a methyl donor. Dithiothreitol protected the sulfhydryl groups in the membrane and caused a concentration- and time-dependent increase of phospholipid N-methylation at three different catalytic sites. This stimulation was highest (9-fold) in the presence of 1 MM MgCl2 and 0.1 μM S-adenosyl-L-[methyl-3H]methionine at pH 8.0 (catalytic site 1), and was associated with an enhancement of Vmax without changes in Km for the methyl donor. Thiol glutathione was less stimulatory than dithiothreitol; glutathione disulfide inhibited the phosphatidylethanolamine N-methylation by 50{\%}. The alkylating reagents, N-ethylmaleimide and methylmethanethiosulfonate, inhibited the N-methylation with IC5O of 6.9 and 14.1 μM, respectively; this inhibition was prevented by 1 mM dithiothreitol. These results indicate a critical role of sulfhydryl groups for the activity of the cardiac sarcolemmal phosphatidylethanolamine N-methyltransferase and suggest that this enzyme system in cardiac sarcolemma may be controlled by the glutathione/glutathione disulfide redox state in the cell.",
keywords = "neutral lipid methylation, phospholipid N-methylation, rat heart, S-adenosyl-L-methionine, sarcolemma, sulfhydryl groups",
author = "Roland Vetter and Jian Dai and Nasrin Mesaeli and Vincenzo Panagia and Dhalla, {Naranjan S.}",
year = "1991",
month = "4",
doi = "10.1007/BF00229596",
language = "English",
volume = "103",
pages = "85--96",
journal = "Molecular and Cellular Biochemistry",
issn = "0300-8177",
publisher = "Springer Netherlands",
number = "1",

}

TY - JOUR

T1 - Role of sulfhydryl groups in phospholipid methylation reactions of cardiac sarcolemma

AU - Vetter, Roland

AU - Dai, Jian

AU - Mesaeli, Nasrin

AU - Panagia, Vincenzo

AU - Dhalla, Naranjan S.

PY - 1991/4

Y1 - 1991/4

N2 - The effect of reagents that modify sulfur-containing amino acid residues in the phosphatidylethanolamine N-methyltransferase was studied in the isolated rat cardiac sarcolemma by employing S-adenosyl-L-[methyl-3H]methionine as a methyl donor. Dithiothreitol protected the sulfhydryl groups in the membrane and caused a concentration- and time-dependent increase of phospholipid N-methylation at three different catalytic sites. This stimulation was highest (9-fold) in the presence of 1 MM MgCl2 and 0.1 μM S-adenosyl-L-[methyl-3H]methionine at pH 8.0 (catalytic site 1), and was associated with an enhancement of Vmax without changes in Km for the methyl donor. Thiol glutathione was less stimulatory than dithiothreitol; glutathione disulfide inhibited the phosphatidylethanolamine N-methylation by 50%. The alkylating reagents, N-ethylmaleimide and methylmethanethiosulfonate, inhibited the N-methylation with IC5O of 6.9 and 14.1 μM, respectively; this inhibition was prevented by 1 mM dithiothreitol. These results indicate a critical role of sulfhydryl groups for the activity of the cardiac sarcolemmal phosphatidylethanolamine N-methyltransferase and suggest that this enzyme system in cardiac sarcolemma may be controlled by the glutathione/glutathione disulfide redox state in the cell.

AB - The effect of reagents that modify sulfur-containing amino acid residues in the phosphatidylethanolamine N-methyltransferase was studied in the isolated rat cardiac sarcolemma by employing S-adenosyl-L-[methyl-3H]methionine as a methyl donor. Dithiothreitol protected the sulfhydryl groups in the membrane and caused a concentration- and time-dependent increase of phospholipid N-methylation at three different catalytic sites. This stimulation was highest (9-fold) in the presence of 1 MM MgCl2 and 0.1 μM S-adenosyl-L-[methyl-3H]methionine at pH 8.0 (catalytic site 1), and was associated with an enhancement of Vmax without changes in Km for the methyl donor. Thiol glutathione was less stimulatory than dithiothreitol; glutathione disulfide inhibited the phosphatidylethanolamine N-methylation by 50%. The alkylating reagents, N-ethylmaleimide and methylmethanethiosulfonate, inhibited the N-methylation with IC5O of 6.9 and 14.1 μM, respectively; this inhibition was prevented by 1 mM dithiothreitol. These results indicate a critical role of sulfhydryl groups for the activity of the cardiac sarcolemmal phosphatidylethanolamine N-methyltransferase and suggest that this enzyme system in cardiac sarcolemma may be controlled by the glutathione/glutathione disulfide redox state in the cell.

KW - neutral lipid methylation

KW - phospholipid N-methylation

KW - rat heart

KW - S-adenosyl-L-methionine

KW - sarcolemma

KW - sulfhydryl groups

UR - http://www.scopus.com/inward/record.url?scp=0025844807&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025844807&partnerID=8YFLogxK

U2 - 10.1007/BF00229596

DO - 10.1007/BF00229596

M3 - Article

VL - 103

SP - 85

EP - 96

JO - Molecular and Cellular Biochemistry

JF - Molecular and Cellular Biochemistry

SN - 0300-8177

IS - 1

ER -