In Vivo Protein Targets Of Reactive Lipid Peroxidation Products

摘要

Reactive secondary end products of lipid peroxidation, such as alpha,beta-unsaturated aldeydes, keto aldehydes and 4-hydroxy-2-alkenals, modify proteins and cause loss of protein function. We have developed chemical approaches for the identification, characterization and quantitation of aldehyde-mediated modifications to proteins associated with oxidative stress: (A) chemical tagging of oxylipid-peptides and proteins using the aldehyde reactive affinity probe N'-aminooxymethylcarbonylhydrazino D-biotin, (B) a pull-down approach using hydrazide-functionalized beads and (C) HICAT, a hydrazide-functionalized isotope-coded affinity tag for the selective isolation and quantitative analysis of proteins modified by alpha,beta-unsaturated aldehydes. We have identified and determined the site of modification of major in vivo protein targets of Michael-type addition reactions caused by alpha,beta-unsaturated aldehydic lipid peroxidation products in rat cardiac mitochondria.