Cardiac nitric oxide synthase-1 localization within the cardiomyocyte is accompanied by the adaptor protein, CAPON

Farideh Beigi, Behzad N. Oskouei, Meizi Zheng, Carol A. Cooke, Guillaume Lamirault, Joshua M. Hare

Research output: Contribution to journalArticle

Abstract

The mechanism(s) regulating nitric oxide synthase-1 (NOS1) localization within the cardiac myocyte in health and disease remains unknown. Here we tested the hypothesis that the PDZ-binding domain interaction between CAPON (carboxy-terminal PDZ ligand of NOS1), a NOS1 adaptor protein and NOS1, contribute to NOS1 localization in specific organelles within cardiomyocytes. Ventricular cardiomyocytes and whole heart homogenates were isolated from sham and post-myocardial infarction (MI) wild-type (C57BL/6) and NOS1-/- female mice for quantification of CAPON protein expression levels. NOS1, CAPON, xanthine oxidoreductase and Dexras1, a CAPON binding partner, were all present and enriched in isolated cardiac sarcoplasmic reticulum (SR) fractions. CAPON co-immunoprecipitated with the mu and alpha isoforms of NOS1 in whole heart lysates, and co-localization of CAPON and NOS1 was demonstrated in the SR and mitochondria with dual immuno-gold electron microscopy. Following MI, CAPON and NOS1 both redistributed to caveolae and colocalized with caveolin-3. In addition, following MI, expression level of CAPON remained unchanged and Dexras1 was reduced, CAPON binding to xanthine oxidoreductase was augmented and the plasma membrane calcium ATPase (PMCA) increased. In NOS1 deficient myocytes, CAPON abundance in the SR was reduced, and redistribution to caveolae and PMCA binding after MI was absent. Together these findings support the hypothesis that NOS1 redistribution in injured myocardium requires the formation of a complex with the PDZ adaptor protein CAPON.

Original languageEnglish (US)
Pages (from-to)226-233
Number of pages8
JournalNitric Oxide - Biology and Chemistry
Volume21
Issue number3-4
DOIs
StatePublished - Dec 15 2009
Externally publishedYes

Fingerprint

Cardiac Myocytes
Nitric Oxide Synthase
Ligands
Proteins
Plasma Membrane Calcium-Transporting ATPases
Sarcoplasmic Reticulum
Myocardial Infarction
Xanthine Dehydrogenase
Caveolae
Caveolin 3
PDZ Domains
Nitric Oxide Synthase Type I
Mitochondria
Immunoelectron Microscopy
Gold
Organelles
Muscle Cells
Electron microscopy
Myocardium
Protein Isoforms

Keywords

  • CAPON
  • Dexras1
  • Heart failure
  • Nitric oxide synthase-1 (NOS1)
  • PDZ-interaction
  • PMCA

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Biochemistry
  • Cancer Research
  • Physiology

Cite this

Cardiac nitric oxide synthase-1 localization within the cardiomyocyte is accompanied by the adaptor protein, CAPON. / Beigi, Farideh; Oskouei, Behzad N.; Zheng, Meizi; Cooke, Carol A.; Lamirault, Guillaume; Hare, Joshua M.

In: Nitric Oxide - Biology and Chemistry, Vol. 21, No. 3-4, 15.12.2009, p. 226-233.

Research output: Contribution to journalArticle

Beigi, Farideh ; Oskouei, Behzad N. ; Zheng, Meizi ; Cooke, Carol A. ; Lamirault, Guillaume ; Hare, Joshua M. / Cardiac nitric oxide synthase-1 localization within the cardiomyocyte is accompanied by the adaptor protein, CAPON. In: Nitric Oxide - Biology and Chemistry. 2009 ; Vol. 21, No. 3-4. pp. 226-233.
@article{0430c5fbaef54e76869852d50b194717,
title = "Cardiac nitric oxide synthase-1 localization within the cardiomyocyte is accompanied by the adaptor protein, CAPON",
abstract = "The mechanism(s) regulating nitric oxide synthase-1 (NOS1) localization within the cardiac myocyte in health and disease remains unknown. Here we tested the hypothesis that the PDZ-binding domain interaction between CAPON (carboxy-terminal PDZ ligand of NOS1), a NOS1 adaptor protein and NOS1, contribute to NOS1 localization in specific organelles within cardiomyocytes. Ventricular cardiomyocytes and whole heart homogenates were isolated from sham and post-myocardial infarction (MI) wild-type (C57BL/6) and NOS1-/- female mice for quantification of CAPON protein expression levels. NOS1, CAPON, xanthine oxidoreductase and Dexras1, a CAPON binding partner, were all present and enriched in isolated cardiac sarcoplasmic reticulum (SR) fractions. CAPON co-immunoprecipitated with the mu and alpha isoforms of NOS1 in whole heart lysates, and co-localization of CAPON and NOS1 was demonstrated in the SR and mitochondria with dual immuno-gold electron microscopy. Following MI, CAPON and NOS1 both redistributed to caveolae and colocalized with caveolin-3. In addition, following MI, expression level of CAPON remained unchanged and Dexras1 was reduced, CAPON binding to xanthine oxidoreductase was augmented and the plasma membrane calcium ATPase (PMCA) increased. In NOS1 deficient myocytes, CAPON abundance in the SR was reduced, and redistribution to caveolae and PMCA binding after MI was absent. Together these findings support the hypothesis that NOS1 redistribution in injured myocardium requires the formation of a complex with the PDZ adaptor protein CAPON.",
keywords = "CAPON, Dexras1, Heart failure, Nitric oxide synthase-1 (NOS1), PDZ-interaction, PMCA",
author = "Farideh Beigi and Oskouei, {Behzad N.} and Meizi Zheng and Cooke, {Carol A.} and Guillaume Lamirault and Hare, {Joshua M.}",
year = "2009",
month = "12",
day = "15",
doi = "10.1016/j.niox.2009.09.005",
language = "English (US)",
volume = "21",
pages = "226--233",
journal = "Nitric Oxide - Biology and Chemistry",
issn = "1089-8603",
publisher = "Academic Press Inc.",
number = "3-4",

}

TY - JOUR

T1 - Cardiac nitric oxide synthase-1 localization within the cardiomyocyte is accompanied by the adaptor protein, CAPON

AU - Beigi, Farideh

AU - Oskouei, Behzad N.

AU - Zheng, Meizi

AU - Cooke, Carol A.

AU - Lamirault, Guillaume

AU - Hare, Joshua M.

PY - 2009/12/15

Y1 - 2009/12/15

N2 - The mechanism(s) regulating nitric oxide synthase-1 (NOS1) localization within the cardiac myocyte in health and disease remains unknown. Here we tested the hypothesis that the PDZ-binding domain interaction between CAPON (carboxy-terminal PDZ ligand of NOS1), a NOS1 adaptor protein and NOS1, contribute to NOS1 localization in specific organelles within cardiomyocytes. Ventricular cardiomyocytes and whole heart homogenates were isolated from sham and post-myocardial infarction (MI) wild-type (C57BL/6) and NOS1-/- female mice for quantification of CAPON protein expression levels. NOS1, CAPON, xanthine oxidoreductase and Dexras1, a CAPON binding partner, were all present and enriched in isolated cardiac sarcoplasmic reticulum (SR) fractions. CAPON co-immunoprecipitated with the mu and alpha isoforms of NOS1 in whole heart lysates, and co-localization of CAPON and NOS1 was demonstrated in the SR and mitochondria with dual immuno-gold electron microscopy. Following MI, CAPON and NOS1 both redistributed to caveolae and colocalized with caveolin-3. In addition, following MI, expression level of CAPON remained unchanged and Dexras1 was reduced, CAPON binding to xanthine oxidoreductase was augmented and the plasma membrane calcium ATPase (PMCA) increased. In NOS1 deficient myocytes, CAPON abundance in the SR was reduced, and redistribution to caveolae and PMCA binding after MI was absent. Together these findings support the hypothesis that NOS1 redistribution in injured myocardium requires the formation of a complex with the PDZ adaptor protein CAPON.

AB - The mechanism(s) regulating nitric oxide synthase-1 (NOS1) localization within the cardiac myocyte in health and disease remains unknown. Here we tested the hypothesis that the PDZ-binding domain interaction between CAPON (carboxy-terminal PDZ ligand of NOS1), a NOS1 adaptor protein and NOS1, contribute to NOS1 localization in specific organelles within cardiomyocytes. Ventricular cardiomyocytes and whole heart homogenates were isolated from sham and post-myocardial infarction (MI) wild-type (C57BL/6) and NOS1-/- female mice for quantification of CAPON protein expression levels. NOS1, CAPON, xanthine oxidoreductase and Dexras1, a CAPON binding partner, were all present and enriched in isolated cardiac sarcoplasmic reticulum (SR) fractions. CAPON co-immunoprecipitated with the mu and alpha isoforms of NOS1 in whole heart lysates, and co-localization of CAPON and NOS1 was demonstrated in the SR and mitochondria with dual immuno-gold electron microscopy. Following MI, CAPON and NOS1 both redistributed to caveolae and colocalized with caveolin-3. In addition, following MI, expression level of CAPON remained unchanged and Dexras1 was reduced, CAPON binding to xanthine oxidoreductase was augmented and the plasma membrane calcium ATPase (PMCA) increased. In NOS1 deficient myocytes, CAPON abundance in the SR was reduced, and redistribution to caveolae and PMCA binding after MI was absent. Together these findings support the hypothesis that NOS1 redistribution in injured myocardium requires the formation of a complex with the PDZ adaptor protein CAPON.

KW - CAPON

KW - Dexras1

KW - Heart failure

KW - Nitric oxide synthase-1 (NOS1)

KW - PDZ-interaction

KW - PMCA

UR - http://www.scopus.com/inward/record.url?scp=71849110674&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=71849110674&partnerID=8YFLogxK

U2 - 10.1016/j.niox.2009.09.005

DO - 10.1016/j.niox.2009.09.005

M3 - Article

C2 - 19800018

AN - SCOPUS:71849110674

VL - 21

SP - 226

EP - 233

JO - Nitric Oxide - Biology and Chemistry

JF - Nitric Oxide - Biology and Chemistry

SN - 1089-8603

IS - 3-4

ER -