Global and Targeted Proteomic Identification of oxidative modifications.
EMSL Project ID
17992
Abstract
Recent research has provided an emerging picture of the significant role of reactive oxygen/nitrogen species (ROS/RNS) and their associated modification of cellular proteins in cellular signaling/regulation as well as in pathology. The EMSL proteomic work we propose here stems from previous work by ourselves and others demonstrating the SR Ca-ATPase in aging heart and skeletal muscle as well as athleroschlerotic aorta incurs nitrotyrosine modifications at specific sites (Tyr294,294) that result in partial inhibition of Ca-ATPase activity with implications for contractile function (Knyushko, 2005; Adachi, 2005). Additional researchers have observed tyrosine nitration in over 80 different pathologies, many associated with misfolding diseases as in neurodegenerative disorders. Recent analysis of endogenously nitrated proteins in mouse brain, enabled by the high sensitivity of EMSL MS capabilities, have shown that the majority of these proteins have been associated with defects in neurodegenerative diseases (Sackstedter, Qian, 2006). Thus, the appearance of these modified proteins in vivo in healthy animals suggests sites of vulnerability where inflammation-derived nitration may contribute to the progression of disease. Therefore, we propose to utilize the mass spect capabilities at EMSL for (1) identification of oxidative modifications related to nitrative stress, e.g., nitrotyrosine and methionine sulfoxides in heart and skeletal muscle and myocyte models of nitrative stress, asking if these may be related to disease; and,(2) continued exploration of the role of these modifications in neurodegenerative disease models and in prion disease(where the scrapie prion protein has recently been found to be nitrated). In addition, (3) we are developing assays for organelle and protein complex isolation in muscle and myocytes to address if oxidative modifications alter protein localization and interactions with the Ca-ATPase and its regulatory partners. Project Details
Project type
Exploratory Research
Start Date
2006-03-02
End Date
2007-08-09
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members