Comparative Proteomics Reveals Dysregulated Mitochondrial O-GlcNAcylation in Diabetic Hearts

J Proteome Res. 2016 Jul 1;15(7):2254-64. doi: 10.1021/acs.jproteome.6b00250. Epub 2016 Jun 2.


O-linked β-N-acetylglucosamine (O-GlcNAc), a post-translational modification on serine and threonine residues of many proteins, plays crucial regulatory roles in diverse biological events. As a nutrient sensor, O-GlcNAc modification (O-GlcNAcylation) on nuclear and cytoplasmic proteins underlies the pathology of diabetic complications including cardiomyopathy. However, mitochondrial O-GlcNAcylation, especially in response to chronic hyperglycemia in diabetes, has been poorly explored. We performed a comparative O-GlcNAc profiling of mitochondria from control and streptozotocin (STZ)-induced diabetic rat hearts by using an improved β-elimination/Michael addition with isotopic DTT reagents (BEMAD) followed by tandem mass spectrometric analysis. In total, 86 mitochondrial proteins, involved in diverse pathways, were O-GlcNAcylated. Among them, many proteins have site-specific alterations in O-GlcNAcylation in response to diabetes, which suggests that protein O-GlcNAcylation is a novel layer of regulation mediating adaptive changes in mitochondrial metabolism during the progression of diabetic cardiomyopathy.

Keywords: O-GlcNAcome; O-GlcNAcylation; diabetic cardiomyopathy; mass spectrometry; mitochondria; proteomics; pyruvate dehydrogenase.

Publication types

  • Comparative Study
  • Research Support, N.I.H., Extramural

MeSH terms

  • Acetylglucosamine / metabolism*
  • Acylation
  • Animals
  • Diabetes Mellitus, Experimental
  • Diabetic Cardiomyopathies / metabolism*
  • Mitochondria / metabolism
  • Mitochondrial Proteins / analysis
  • Mitochondrial Proteins / metabolism*
  • Myocardium / metabolism
  • Proteomics / methods*
  • Rats
  • Tandem Mass Spectrometry


  • Mitochondrial Proteins
  • Acetylglucosamine