BiologicalProteinNitrationMechanismsandSignificance.pptVIP

Protein Nitration Society For Free Radical Biology and Medicine Ischiropoulos * Biological Protein Nitration: Mechanisms and Significance Harry Ischiropoulos Stokes Research Institute, Children’s Hospital of Philadelphia, Department of Biochemistry and Biophysics, The University of Pennsylvania 416D Abramson Center, 34th Street and Civic Center Blvd. Philadelphia, PA 19104-4318 Tel: (215) 590-5320, Fax: (215) 590-4267 email: ischirop@mail.med.upenn.edu The Virtual Free Radical School Post-translational Modifications of Protein Tyrosine Residues Phosphotyrosine Sulfatyrosine Iodotyrosine Dityrosine 3-Nitrotyrosine 3-Chlorotyrosine Biological Protein Nitration: Outline In search of the in vivo nitrating agents Detection of 3-nitrotyrosine and nitrated proteins Specific protein targets Consequences in protein function and in the pathogenesis of disease Metabolism of nitrated proteins Possible role in signal transduction - comparison with other tyrosine signaling pathways and with S-nitrosocysteine Oxidation state (n) 2 3 4 5 Element .NO .NO2 ONOO- Intermediates/ catalysts HNO2 H2O2, HOCl MPO/EPO Hemeproteins Tyr. CO2 Men+ ROH, RCO2 MPO In search of the in vivo nitrating agents: Possible nitration pathways Tyrosyl Radical/?NO: Prostaglandin H Synthase-2, Ribonucleotide Reductase Peroxidases: Catalysts of both nitrite and peroxynitrite-mediated nitration: In vivo contribution has been confirmed by the use of MPO or EPO knock-out mice Hypochlorous acid/NO2- : Likely not involved in peroxidase-mediated nitration Nitrogen Dioxide: Inefficient in the absence of tyrosyl radical ONO(O)CO2- : More efficient nitrating agent than ONOO- in some but not all proteins Tyr. NO2- Protein Nitration Society For Free Radical Biology and Medicine Ischiropoulos 4 Methods for Quantification and Detection of Nitrotyrosine Ana