Muscarinic 2 receptors modulate cardiac proteasome function in a protein kinase G-dependent manner

Mark J. Ranek, Curtis K. Kost, Chengjun Hu, Douglas S. Martin, Xuejun Wang

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Proteasome function insufficiency and inadequate protein quality control are strongly implicated in a large subset of cardiovascular disease and may play an important role in their pathogenesis. Protein degradation by the ubiquitin proteasome system can be physiologically regulated. Cardiac muscarinic 2 (M2) receptors were pharmacologically interrogated in intact mice and cultured neonatal rat ventricular myocytes (NRVMs). Proteasome-mediated proteolysis was measured with a surrogate misfolded protein, proteasome peptidase assay, and by characterizing key proteasome subunits. Successful M2 receptor manipulation in cardiomyocytes was determined by measuring an endogenous protein substrate, and in mice, the cardiovascular physiological response. M2 receptor stimulation was associated with increased proteasome-mediated proteolysis and enhanced peptidase activities, while M2 receptor inhibition yielded opposing results. Additionally, M2 receptor manipulation did not alter abundance of the key proteasome subunits, Rpt6 and β5, but significantly shifted their isoelectric points. Inhibition of protein kinase G abrogated the stimulatory effects on proteasome-mediated proteolysis from M2 receptor activation. We conclude that M2 receptor stimulation enhances, whereas M2 receptor inhibition reduces, proteasome-mediated proteolysis likely through posttranslational modifications. Protein kinase G appears to be the mediator of the M2 receptors actions.

Original languageEnglish (US)
Pages (from-to)43-51
Number of pages9
JournalJournal of Molecular and Cellular Cardiology
Volume69
DOIs
StatePublished - Apr 2014

Fingerprint

Cyclic GMP-Dependent Protein Kinases
Muscarinic Receptors
Proteasome Endopeptidase Complex
Proteolysis
Peptide Hydrolases
Proteins
Isoelectric Point
Post Translational Protein Processing
Ubiquitin
Cardiac Myocytes
Quality Control
Muscle Cells
Cardiovascular Diseases

Keywords

  • Cardiomyocyte
  • Muscarinic 2 receptor
  • Proteasome
  • Protein kinase G

ASJC Scopus subject areas

  • Molecular Biology
  • Cardiology and Cardiovascular Medicine

Cite this

Muscarinic 2 receptors modulate cardiac proteasome function in a protein kinase G-dependent manner. / Ranek, Mark J.; Kost, Curtis K.; Hu, Chengjun; Martin, Douglas S.; Wang, Xuejun.

In: Journal of Molecular and Cellular Cardiology, Vol. 69, 04.2014, p. 43-51.

Research output: Contribution to journalArticle

Ranek, Mark J. ; Kost, Curtis K. ; Hu, Chengjun ; Martin, Douglas S. ; Wang, Xuejun. / Muscarinic 2 receptors modulate cardiac proteasome function in a protein kinase G-dependent manner. In: Journal of Molecular and Cellular Cardiology. 2014 ; Vol. 69. pp. 43-51.
@article{571cfee98328483d875c13a94ecf9ca5,
title = "Muscarinic 2 receptors modulate cardiac proteasome function in a protein kinase G-dependent manner",
abstract = "Proteasome function insufficiency and inadequate protein quality control are strongly implicated in a large subset of cardiovascular disease and may play an important role in their pathogenesis. Protein degradation by the ubiquitin proteasome system can be physiologically regulated. Cardiac muscarinic 2 (M2) receptors were pharmacologically interrogated in intact mice and cultured neonatal rat ventricular myocytes (NRVMs). Proteasome-mediated proteolysis was measured with a surrogate misfolded protein, proteasome peptidase assay, and by characterizing key proteasome subunits. Successful M2 receptor manipulation in cardiomyocytes was determined by measuring an endogenous protein substrate, and in mice, the cardiovascular physiological response. M2 receptor stimulation was associated with increased proteasome-mediated proteolysis and enhanced peptidase activities, while M2 receptor inhibition yielded opposing results. Additionally, M2 receptor manipulation did not alter abundance of the key proteasome subunits, Rpt6 and β5, but significantly shifted their isoelectric points. Inhibition of protein kinase G abrogated the stimulatory effects on proteasome-mediated proteolysis from M2 receptor activation. We conclude that M2 receptor stimulation enhances, whereas M2 receptor inhibition reduces, proteasome-mediated proteolysis likely through posttranslational modifications. Protein kinase G appears to be the mediator of the M2 receptors actions.",
keywords = "Cardiomyocyte, Muscarinic 2 receptor, Proteasome, Protein kinase G",
author = "Ranek, {Mark J.} and Kost, {Curtis K.} and Chengjun Hu and Martin, {Douglas S.} and Xuejun Wang",
year = "2014",
month = "4",
doi = "10.1016/j.yjmcc.2014.01.017",
language = "English (US)",
volume = "69",
pages = "43--51",
journal = "Journal of Molecular and Cellular Cardiology",
issn = "0022-2828",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Muscarinic 2 receptors modulate cardiac proteasome function in a protein kinase G-dependent manner

AU - Ranek, Mark J.

AU - Kost, Curtis K.

AU - Hu, Chengjun

AU - Martin, Douglas S.

AU - Wang, Xuejun

PY - 2014/4

Y1 - 2014/4

N2 - Proteasome function insufficiency and inadequate protein quality control are strongly implicated in a large subset of cardiovascular disease and may play an important role in their pathogenesis. Protein degradation by the ubiquitin proteasome system can be physiologically regulated. Cardiac muscarinic 2 (M2) receptors were pharmacologically interrogated in intact mice and cultured neonatal rat ventricular myocytes (NRVMs). Proteasome-mediated proteolysis was measured with a surrogate misfolded protein, proteasome peptidase assay, and by characterizing key proteasome subunits. Successful M2 receptor manipulation in cardiomyocytes was determined by measuring an endogenous protein substrate, and in mice, the cardiovascular physiological response. M2 receptor stimulation was associated with increased proteasome-mediated proteolysis and enhanced peptidase activities, while M2 receptor inhibition yielded opposing results. Additionally, M2 receptor manipulation did not alter abundance of the key proteasome subunits, Rpt6 and β5, but significantly shifted their isoelectric points. Inhibition of protein kinase G abrogated the stimulatory effects on proteasome-mediated proteolysis from M2 receptor activation. We conclude that M2 receptor stimulation enhances, whereas M2 receptor inhibition reduces, proteasome-mediated proteolysis likely through posttranslational modifications. Protein kinase G appears to be the mediator of the M2 receptors actions.

AB - Proteasome function insufficiency and inadequate protein quality control are strongly implicated in a large subset of cardiovascular disease and may play an important role in their pathogenesis. Protein degradation by the ubiquitin proteasome system can be physiologically regulated. Cardiac muscarinic 2 (M2) receptors were pharmacologically interrogated in intact mice and cultured neonatal rat ventricular myocytes (NRVMs). Proteasome-mediated proteolysis was measured with a surrogate misfolded protein, proteasome peptidase assay, and by characterizing key proteasome subunits. Successful M2 receptor manipulation in cardiomyocytes was determined by measuring an endogenous protein substrate, and in mice, the cardiovascular physiological response. M2 receptor stimulation was associated with increased proteasome-mediated proteolysis and enhanced peptidase activities, while M2 receptor inhibition yielded opposing results. Additionally, M2 receptor manipulation did not alter abundance of the key proteasome subunits, Rpt6 and β5, but significantly shifted their isoelectric points. Inhibition of protein kinase G abrogated the stimulatory effects on proteasome-mediated proteolysis from M2 receptor activation. We conclude that M2 receptor stimulation enhances, whereas M2 receptor inhibition reduces, proteasome-mediated proteolysis likely through posttranslational modifications. Protein kinase G appears to be the mediator of the M2 receptors actions.

KW - Cardiomyocyte

KW - Muscarinic 2 receptor

KW - Proteasome

KW - Protein kinase G

UR - http://www.scopus.com/inward/record.url?scp=84895461701&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84895461701&partnerID=8YFLogxK

U2 - 10.1016/j.yjmcc.2014.01.017

DO - 10.1016/j.yjmcc.2014.01.017

M3 - Article

C2 - 24508699

AN - SCOPUS:84895461701

VL - 69

SP - 43

EP - 51

JO - Journal of Molecular and Cellular Cardiology

JF - Journal of Molecular and Cellular Cardiology

SN - 0022-2828

ER -