title: Skeletal muscle aldolase an overexpression in endotoxemic rats and inhibited by GSNO via potential role for S-nitrosylation in vitro.
abstract: BACKGROUND
  Hepatic aldolase (ALD) A mRNA transcription and ALD B S-nitrosylation have been confirmed in endotoxemic rats and mice, respectively. In the present study we investigated whether the skeletal muscle ALD A shared potential for S-nitrosylation to act as a hypoxia-related signaling mechanism in lipopolysaccharide (LPS) challenged rats.
  
  
  METHODS
  Male Sprague Dawley rats were treated (i.p.) as follows, control group (n = 6) with 0.9% NaCl, tested group (n = 6) with a single dose of 2 mg/kg LPS. Protein S-nitrosylation was determined by biotin switch and dot blotting analysis. ALD A, hypoxia-inducible factor 1α and vascular endothelial growth factor were determined by western blotting. ALD A catalytic activity treated with S-nitrosoglutathione (GSNO), an exogenous NO-donor, was examined in vitro.
  
  
  RESULTS
  There were several S-nitrosylated proteins under basal conditions. ALD A was over-expressed in a hypoxia-related way in the skeletal muscle of LPS challenged rats. Importantly, treatment of ALD A with GSNO at concentration 50 μmol/L ∼ 1000 μmol/L that inhibited catalytic activity, increased the number of S-nitrosylated bands and led to hyper-nitrosylation of basally S-nitrosylated proteins of ALD A. Quantization of enzyme S-nitrosothiol showed that a maximal of four cysteines per subunit was modified by S-nitrosylation in the presence of GSNO.
  
  
  CONCLUSIONS
  These findings suggested that S-nitrosylation of ALD A might serve as a novel mechanism for controlling ALD A activity at the post-translational level in endotoxemic rats.
authors.affiliation: Zhejiang-California International NanoSystems Institute, Zhejiang University, Hangzhou, China.
authors.forename: Jieping
authors.initials: J
authors.lastname: Yan
authors.forename: Qiang
authors.initials: Q
authors.lastname: Shi
authors.forename: Zhe
authors.initials: Z
authors.lastname: Chen
authors.forename: Renyao
authors.initials: R
authors.lastname: Zhuang
authors.forename: Haifei
authors.initials: H
authors.lastname: Chen
authors.forename: Danyan
authors.initials: D
authors.lastname: Zhu
authors.forename: Yijia
authors.initials: Y
authors.lastname: Lou
mesh_terms: D000818:Animals; D019446:Endotoxemia / Q000201:enzymology*; D005634:Fructose-Bisphosphate Aldolase / Q000037:antagonists & inhibitors / Q000502:physiology*; D051795:Hypoxia-Inducible Factor 1, alpha Subunit / Q000032:analysis; D008070:Lipopolysaccharides / Q000633:toxicity; D008297:Male; D018482:Muscle, Skeletal / Q000201:enzymology*; D011499:Protein Processing, Post-Translational; D051381:Rats; D017207:Rats, Sprague-Dawley; D026422:S-Nitrosoglutathione / Q000494:pharmacology*; D042461:Vascular Endothelial Growth Factor A / Q000032:analysis
chemical_list: C497444:Hif1a protein, rat; D051795:Hypoxia-Inducible Factor 1, alpha Subunit; D008070:Lipopolysaccharides; D042461:Vascular Endothelial Growth Factor A; D026422:S-Nitrosoglutathione; D005634:Fructose-Bisphosphate Aldolase