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. Free Radic Biol Med 2009 Feb 15;46(4):454-61
Date
12/04/2008Pubmed ID
19049863Pubmed Central ID
PMC2661569DOI
10.1016/j.freeradbiomed.2008.10.046Scopus ID
2-s2.0-58649090242 (requires institutional sign-in at Scopus site) 31 CitationsAbstract
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.
Author List
Chatterjee S, Ehrenshaft M, Bhattacharjee S, Deterding LJ, Bonini MG, Corbett J, Kadiiska MB, Tomer KB, Mason RPMESH terms used to index this publication - Major topics in bold
AllopurinolAmidines
Animals
Benzylamines
Carboxypeptidase B
Cross-Linking Reagents
Cyclic N-Oxides
Electron Spin Resonance Spectroscopy
Free Radicals
Immunoprecipitation
Lipopolysaccharides
Male
Mass Spectrometry
Mice
Mice, Inbred C57BL
Mice, Knockout
Nitric Oxide Synthase Type III
Protein Processing, Post-Translational
Shock, Septic
Spleen
Xanthine Oxidase
