Immuno-spin trapping of a post-translational carboxypeptidase B1 radical formed by a dual role of xanthine oxidase and endothelial nitric oxide synthase in acute septic mice

Saurabh Chatterjee*, Marilyn Ehrenshaft, Suchandra Bhattacharjee, Leesa J. Deterding, Marcelo G. Bonini, Jean Corbett, Maria B. Kadiiska, Kenneth B. Tomer, Ronald P. Mason

*Corresponding author for this work

Research output: Contribution to journalArticle

29 Scopus citations

Abstract

Post-translational modification of proteins due to exposure to radicals and other reactive species are markers of metabolic and inflammatory oxidative stress such as sepsis. This study uses the nitrone spin-trap DMPO and a combination of immuno-spin trapping and mass spectrometry to identify in vivo products of radical reactions in mice. We report the detection of dose-dependent production of DMPO-carboxypeptidase B1 (CPB1) adducts in the spleens of mice treated with lipopolysaccharide (LPS). Additionally, we report significant detection of DMPO-CPB1 adducts in mice experiencing normal physiological conditions. Treatments with inhibitors and experiments with knock-out mice indicate that xanthine oxidase and endothelial nitric oxide synthase are important sources of the reactive species that lead to CPB1 adduct formation. We also report a significant loss of CPB1 activity following LPS challenge in conjunction with an increase in CPB1 protein accumulation. This suggests the presence of a possible mechanism for CPB1 activity loss with compensatory protein production.

Original languageEnglish (US)
Pages (from-to)454-461
Number of pages8
JournalFree Radical Biology and Medicine
Volume46
Issue number4
DOIs
StatePublished - Feb 15 2009
Externally publishedYes

Keywords

  • Immuno-spin trapping
  • Inflammation
  • Nitrone adduct
  • Oxidative stress

ASJC Scopus subject areas

  • Biochemistry
  • Physiology (medical)

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