Immuno-spin trapping from biochemistry to medicine: Advances,challenges, and pitfalls. Focus on protein-centered radicals |
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Authors: | Sandra E Gomez-Mejiba Zili Zhai Maria C Della-Vedova Marcos D Muñoz Saurabh Chatterjee Rheal A Towner Kenneth Hensley Robert A Floyd Ronald P Mason Dario C Ramirez |
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Institution: | 1. Laboratory of Experimental Medicine and Therapeutics, Institute Multidisciplinary of Biological Investigations-San Luis (IMIBIO-SL), National Bureau of Science and Technology (CONICET) and National University of San Luis, San Luis, 5700 San Luis, Argentina;2. Section of Gastroenterology, Department of Gastroenterology, The University of Chicago, IL 60637, USA;3. Environmental Health & Disease Laboratory, Department of Environmental Health Sciences, University of South Carolina, Columbia, 29208 SC, USA;4. Advanced Magnetic Resonance Center, Oklahoma Medical Research Foundation, Oklahoma City, 73104 OK, USA;5. Department of Pathology, University of Toledo Medical Center, Toledo, 43614 OH, USA;6. Laboratory of Experimental Therapeutics, Oklahoma Medical Research Foundation, Oklahoma City, 74104 OK, USA;g Laboratory of Pharmacology and Toxicology, National Institute of Environmental Health Sciences, Research Triangle Park, 23709 NC, USA;h Molecular Physiology, Molecular Biology Section, Department of Biochemistry and Biological Sciences, National University of San Luis, 5700 San Luis, San Luis, Argentina |
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Abstract: | BackgroundImmuno-spin trapping (IST) is based on the reaction of a spin trap with a free radical to form a stable nitrone adduct, followed by the use of antibodies, rather than traditional electron paramagnetic resonance spectroscopy, to detect the nitrone adduct. IST has been successfully applied to mechanistic in vitro studies, and recently, macromolecule-centered radicals have been detected in models of drug-induced agranulocytosis, hepatotoxicity, cardiotoxicity, and ischemia/reperfusion, as well as in models of neurological, metabolic and immunological diseases.Scope of the reviewTo critically evaluate advances, challenges, and pitfalls as well as the scientific opportunities of IST as applied to the study of protein-centered free radicals generated in stressed organelles, cells, tissues and animal models of disease and exposure.Major conclusionsBecause the spin trap has to be present at high enough concentrations in the microenvironment where the radical is formed, the possible effects of the spin trap on gene expression, metabolism and cell physiology have to be considered in the use of IST and in the interpretation of results. These factors have not yet been thoroughly dealt with in the literature.General significanceThe identification of radicalized proteins during cell/tissue response to stressors will help define their role in the complex cellular response to stressors and pathogenesis; however, the fidelity of spin trapping/immuno-detection and the effects of the spin trap on the biological system should be considered.This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn. |
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Keywords: | AG aminoglutethimide BSA bovine serum albumin DMPO 5 5-dimethyl-1-pyrroline N-oxide ESR electron spin resonance HPLC high performance liquid chromatography hoMb horse myglobin huHb/Mb human hemoglobin/myoglobin I/R ischemia/reperfusion IST immuno-spin trapping LC liquid chromatography LPS lipopolysaccharide MRI magnetic resonance imaging MS mass spectrometry OA octanoic acid SOD superoxide dismutase TPO thyroid peroxidase |
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