Emerging evidence from several laboratories indicates that lipid oxidation products make major contributionsto human disease. An important class of lipid oxidation products are electrophiles that react with cellularnucleophiles including glutathione, proteins, and DNA. Although glutathione and DMA modification has beenstudied for many years, inadequate technology has limited our ability to investigate protein modification andits relation to biological outcome. Technological advances in proteomics and genomics now make it possibleto assemble comprehensive profiles of proteins modified by lipid electrophiles as well as the transcriptionalchanges they induce. Comparison of these dense datasets using systems approaches enables hypothesesto be generated linking protein modification to cellular response. Research in the Program Project over thepast two years has generated critical reagents and novel approaches for identifying lipid electrophilemodifiedproteins and their biological consequences. We are now poised to mount an aggressive campaignto define the chemistry and biology of protein modification by lipid electrophiles and its relationship toinflammation, a major contributor to many human diseases. The role of Project 3 is to identify the mostbiologically interesting lipid electrophiles emanating from research in Projects 1 and.2 and to define thetranscriptional changes that they induce in the human macrophage cell line, THP-1. This information will becompared to the pattern of protein modification by the same agents, which will be determined in Project 4,and hypotheses formulated linking protein modification to biological outcome. These hypotheses will betested using a combination of approaches such as RNA knockdown, ectopic protein expression, andbiochemical assays. This will enable the elucidation of mechanisms by which protein modifications by lipidlectrophiles lead to pro- and anti-inflammatory responses.
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