Skeletal Muscle Aldolase an Overexpression in Endotoxemic Rats and Inhibited by Gsno Via Potential Role for S-Nitrosylation in Vitro

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. MATERIALS AND 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.

文章引用产品

• 

  • 
  • GAR007
  • 二抗

  • Goat Anti-Rabbit IgG(H+L) HRP 山羊抗兔二抗-HRP

  • ¥289.00 – ¥350.00