O-linked GlcNAc modification of cardiac myofilament proteins

A novel regulator of myocardial contractile function

Genaro A. Ramirez-Correa, Wenhai Jin, Zihao Wang, Xin Zhong, Wei Dong Gao, Wagner B. Dias, Cecilia Vecoli, Gerald Warren Hart, Anne M Murphy

Research output: Contribution to journalArticle

Abstract

In addition to O-phosphorylation, O-linked modifications of serine and threonine by β-N-acetyl-d-glucosamine (GlcNAc) may regulate muscle contractile function. This study assessed the potential role of O-GlcNAcylation in cardiac muscle contractile activation. To identify specific sites of O-GlcNAcylation in cardiac myofilament proteins, a recently developed methodology based on GalNAz-biotin labeling followed by dithiothreitol replacement and light chromatography/tandem mass spectrometry site mapping was adopted. Thirty-two O-GlcNAcylated peptides from cardiac myofilaments were identified on cardiac myosin heavy chain, actin, myosin light chains, and troponin I. To assess the potential physiological role of the GlcNAc, force-[Ca] relationships were studied in skinned rat trabeculae. Exposure to GlcNAc significantly decreased calcium sensitivity (pCa50), whereas maximal force (Fmax) and Hill coefficient (n) were not modified. Using a pan-specific O-GlcNAc antibody, it was determined that acute exposure of myofilaments to GlcNAc induced a significant increase in actin O-GlcNAcylation. This study provides the first identification of O-GlcNAcylation sites in cardiac myofilament proteins and demonstrates their potential role in regulating myocardial contractile function.

Original languageEnglish (US)
Pages (from-to)1354-1358
Number of pages5
JournalCirculation Research
Volume103
Issue number12
DOIs
StatePublished - Dec 5 2008

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Myofibrils
Staphylococcal Protein A
Actins
Cardiac Myosins
Troponin I
Myosin Heavy Chains
Dithiothreitol
Glucosamine
Threonine
Biotin
Tandem Mass Spectrometry
Serine
Chromatography
Myocardium
Proteins
Phosphorylation
Calcium
Light
Muscles
Peptides

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

O-linked GlcNAc modification of cardiac myofilament proteins : A novel regulator of myocardial contractile function. / Ramirez-Correa, Genaro A.; Jin, Wenhai; Wang, Zihao; Zhong, Xin; Gao, Wei Dong; Dias, Wagner B.; Vecoli, Cecilia; Hart, Gerald Warren; Murphy, Anne M.

In: Circulation Research, Vol. 103, No. 12, 05.12.2008, p. 1354-1358.

Research output: Contribution to journalArticle

Ramirez-Correa, Genaro A. ; Jin, Wenhai ; Wang, Zihao ; Zhong, Xin ; Gao, Wei Dong ; Dias, Wagner B. ; Vecoli, Cecilia ; Hart, Gerald Warren ; Murphy, Anne M. / O-linked GlcNAc modification of cardiac myofilament proteins : A novel regulator of myocardial contractile function. In: Circulation Research. 2008 ; Vol. 103, No. 12. pp. 1354-1358.
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