title: Genetically encoding protein oxidative damage.
abstract: Posttranslational modification of tyrosine residues in proteins, to produce 3-nitrotyrosine (3-NT), is associated with over 50 disease states including transplant rejection, lung infection, central nervous system and ocular inflammation shock, cancer, and neurological disorders (for example, Alzheimer's disease, Parkinson's disease, and stroke). The levels of 3-NT increase in aging tissue, and levels of 3-NT in proteins are a predictor of disease risk. Here we report the evolution and characterization of an aminoacyl-tRNA synthetase/tRNA pair for the cotranslational, site-specific incorporation of 3-NT into proteins at genetically encoded sites. To demonstrate the utility of our approach for studying the effect on protein function of nitration on sites defined in vivo, we prepared manganese superoxide dismutase (MnSOD) that is homogeneously nitrated at a site known to be modified in disease-related inflammatory responses, and we measured the effect of this defined modification on protein function.
authors.affiliation: Medical Research Council Laboratory of Molecular Biology, Hills Road, Cambridge CB2 0QH, United Kingdom.
authors.forename: Heinz
authors.initials: H
authors.lastname: Neumann
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Hazen
authors.forename: John
authors.initials: J
authors.lastname: Weinstein
authors.forename: Ryan A
authors.initials: RA
authors.lastname: Mehl
authors.forename: Jason W
authors.initials: JW
authors.lastname: Chin
mesh_terms: D000604:Amino Acyl-tRNA Synthetases / Q000737:chemistry* / Q000235:genetics; D001665:Binding Sites; D004926:Escherichia coli / Q000235:genetics; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D010084:Oxidation-Reduction; D012343:RNA, Transfer / Q000737:chemistry; D013482:Superoxide Dismutase / Q000737:chemistry*; D014443:Tyrosine / Q000031:analogs & derivatives / Q000737:chemistry
chemical_list: C002744:3-nitrotyrosine; D014443:Tyrosine; D012343:RNA, Transfer; D013482:Superoxide Dismutase; D000604:Amino Acyl-tRNA Synthetases