title: Identification of nitration sites by peroxynitrite on p16 protein.
abstract: Injury of p16 has been implicated in some cancers. In this paper, we focus on the need for identification of peroxynitrite-dependent nitration sites on p16 with HPLC-MS/MS method. Two mono-nitrated residues Tyr129 and Tyr44 were detected in the course of p16 modification induced by peroxynitrite at relative low doses. As suggested by peptide mapping sequence analysis, Tyr44 was more liable to be nitrated by ONOO(-). Study on the chemical environment of two Tyr residues reveals that steric hindrance may be the structural determinant for the nitration sequence. Through technique of SDS-PAGE, ONOO(-) could induce p16 nitration, even strongly damage the combination of p16 with CDK4, which further influence p16's activity.
authors.affiliation: College of Life Science and Bioengineering, Beijing University of Technology, No. 100 Ping Le Yuan, Chaoyang District, Beijing, 100124, People's Republic of China. luoyj@bjut.edu.cn
authors.forename: Yunjing
authors.initials: Y
authors.lastname: Luo
authors.forename: Jingjing
authors.initials: J
authors.lastname: Li
authors.forename: Na
authors.initials: N
authors.lastname: Zhang
authors.forename: Yunhai
authors.initials: Y
authors.lastname: Wang
authors.forename: Rugang
authors.initials: R
authors.lastname: Zhong
mesh_terms: D000595:Amino Acid Sequence; D002851:Chromatography, High Pressure Liquid; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000737:chemistry* / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D009566:Nitrates / Q000032:analysis*; D030421:Peroxynitrous Acid / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D020539:Sequence Analysis, Protein; D053719:Tandem Mass Spectrometry; D014443:Tyrosine / Q000032:analysis*
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D009566:Nitrates; D011994:Recombinant Proteins; D030421:Peroxynitrous Acid; D014443:Tyrosine