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(The FASEB Journal. 2008;22:750.16.)
© 2008 FASEB

750.16

Effect of ischemia-reperfusion on O-GlcNAcylation of specific proteins in isolated rat hearts

Boglarka Laczy1, Landon Wilson2, Charlye A Brocks1, Richard B Marchase3 and John C Chatham1,3

1 Department of Medicine
2 Comprehensive Cancer Center
3 Department of Cell Biology, University of Alabama at Birmingham, Birmingham, AL

ABSTRACT

We have shown that increasing the level of O-linked-N-acetylglucosamine (O-GlcNAc) on proteins protects the heart against ischemia-reperfusion injury; however, the specific proteins altered by O-GlcNAc have yet to be determined. Isolated perfused hearts from male rats (n=5–9/groups) were subjected to 10 min no-flow ischemia (ISC) or ISC followed by 60 min reperfusion (I/R). Overall protein O-GlcNAc levels, determined by anti-O-GlcNAc immunoblots, were increased following ISC (p<0.05) compared to normoxic controls. Using MALDI-TOF we identified proteins in two bands in which O-GlcNAc levels significantly increased during ISC and subsequently decreased following I/R and one band that increased only following I/R. The proteins in the O-GlcNAc positive bands that increased during ISC were identified as glycogen phosphorylase b and mitochondrial aconitase. The protein in the O-GlcNAc band that increased with I/R was identified as the cytoskeletal protein vinculin. Vinculin and aconitase were subsequently immunoprecipitated and both were found to be O-GlcNAc positive by immunoblot analysis. Further studies are needed to determine the effect of O-GlcNAc on the function of these proteins and how this influences the response to I/R. NIH grants: HL067464 and HL079364.





This Article
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