Proteomic identification of 3-nitrotyrosine-containing rat cardiac proteins: effects of biological aging

Abstract

doi:10.1152/ajpheart.01030.2003.—Proteomic techniques were used to identify cardiac proteins from whole heart homogenate and heart mitochondria of Fisher 344/Brown Norway F1 rats, which suffer protein nitration as a consequence of biological aging. Soluble pro-teins from young (5 mo old) and old (26 mo old) animals were separated by one- and two-dimensional gel electrophoresis. One- and two-dimensional Western blots with an anti-nitrotyrosine antibody show an age-related increase in the immunoresponse of a few specific proteins, which were identified by nanoelectrospray ionization-tan-dem mass spectrometry (NSI-MS/MS). Complementary proteins were immunoprecipitated with an immobilized anti-nitrotyrosine antibody followed by NSI-MS/MS analysis. A total of 48 proteins were putatively identified. Among the identified proteins were -enolase, -aldolase, desmin, aconitate hydratase, methylmalonate semialde-hyde dehydrogenase, 3-ketoacyl-CoA thiolase, acetyl-CoA acetyl