Measurement of S-nitrosylated Proteins in Tissues of Rats Fed Diets with Differing Levels of Nitrite
Abstract
Increased ventilation is a general response to shock that may help to maintain cardiac output by improving venous return to the heart. The central stimulus for this increase in ventilation involves the production of S-nitroso-L-cysteine from more ubiquitous precursor S-nitrothiols in the medulla of the brain. Changes in dietary nitrite and nitrate have been shown to increase tissue levels of S-nitrosothiols and to improve survival in models of myocardial infarction. We hypothesize that increased dietary intake of nitrite or nitrate will enhance the ventilatory response in hemorrhage and thereby increase the time of survival. The goal of this project was to adequate methodology for the detection of S-nitrosylated proteins in tissues. A sensitive and specific method for the detection of S-nitrosylated proteins in tissues based on photolysis/immuno-spin-trapping analysis of DMPO thioethers has been developed. The applicability of the method for the quantitative assessments of Snitrosylated proteins in tissues has been tested and confirmed. The ability of dihydrolipoic acid and lipoic acid (LA) plus lipoamide dehydrogenase and NADH to denitrosate S-nitrosocaspase 3 (CASP-SNO) and regulate apoptosis in HepG2 cells has been established.
Document Details
- Document Type
- Technical Report
- Publication Date
- Dec 01, 2011
- Accession Number
- ADA554290
Entities
People
- Valerian E. Kagan
Organizations
- University of Pittsburgh