Hydroxynonenal induces glutathione synthesis through JNK

羟基壬烯醛通过 JNK 诱导谷胱甘肽合成

基本信息

批准号：
7422483
负责人：
HENRY Jay FORMAN
金额：
$ 0.52万
依托单位：
UNIVERSITY OF CALIFORNIA, MERCED
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-09-22 至 2011-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7422483
关键词：
4 hydroxynonenal Address Affinity Chromatography Air Pollutants Anabolism Antibodies Antioxidants Be++ element Beryllium Binding Biological Assay Catalytic Domain Cell Extracts Cells Chromatin Complex Condition DNA Dominant-Negative Mutation Elements Enzyme Induction Enzymes Epithelial Cells Excision Exposure to FarGo GCLC gene GCLM gene Gene Expression Genes Genetic Transcription Glutamate-Cysteine Ligase Glutathione Goals Hour Human Hydrogen Peroxide Immunoprecipitation In Situ Indium Inflammation Injury Interphase Cell Investigation Ligase Gene Lipid Peroxidation Luciferases Lung Lung diseases MAPK14 gene MAPK8 gene Mass Spectrum Analysis Measurement Measures Membrane Metabolism Nuclear Protein Nuclear Proteins Oxidants Oxidative Stress Pathologic Processes Pathway interactions Peptides Phosphotransferases Physiological Post-Translational Protein Processing Protein Overexpression Proteins Range Rate Regulation Reporter Research Personnel Rest Reverse Transcriptase Polymerase Chain Reaction Role Signal Pathway Signal Transduction Signaling Protein Stress System Tissues Transcription Factor AP-1 Transcription factor genes Western Blotting Xenobiotics chromatin immunoprecipitation design human GCLC protein inhibitor/antagonist lung injury oxidation programs protein folding response stress activated protein kinase transcription factor

项目摘要

DESCRIPTION (provided by applicant): This competing renewal application focuses on induction of glutamate cysteine ligase (GCL), the rate limiting enzyme in glutathione (GSH) synthesis, by the lipid peroxidation product, 4-hydroxy-2-nonenal (HNE) through activation of EpRE and TRE cis elements and the JNK signaling pathway. A common response to sublethal exposure to oxidants and electrophiles is an increase in GSH concentration, which is critical to adaptation to greater stress. This increase is dependent on the increased expression of the catalytic and modulatory subunits of glutamate cysteine ligase (GCL, also called (-glutamylcysteine synthetase), the first enzyme in de novo GSH synthesis. HNE in healthy human plasma is 0.3-0.7 micromolar, but increases more than tenfold in inflammatory diseases and exposure to air pollutants. Although HNE can be toxic, at concentrations found in tissues under oxidative stress, it is a potent inducer of GSH, both GCL subunit genes, and other Phase II enzymes. We recently found that transcription factor switching appears to regulate GCL and that inhibition of JNK by a permeable dominant negative peptide completely inhibited GCL induction by HNE while neither ERK or p38MAPK inhibitors were effective. Our hypothesis is that induction of both human GCL genes by HNE occurs through alteration of the compositions of the EpRE and AP-1 binding complexes from those that repress to those that activate transcription. We also hypothesize that the alteration of composition of the EpRE and AP-1 binding complexes is mediated through HNE activation of the JNK signaling pathway. Using human bronchial epithelial cells (HBE1) for all studies, the specific aims of this proposal are to: 1- determine the composition of the complexes binding to the EpRE that regulate GCLC, GCLM, NQO1, and GST4A4 in response to HNE; 2- determine the mechanism of HNE by which HNE causes changing composition of the EpRE binding complexes; 3- determine how the changes in AP-1 complexes in response to HNE relate to the increase of GCL gene transcription; and 4- determine how HNE activates the JNK pathway. The approaches include DNA affinity chromatography, HPLC tandem mass spectrometry, immunodepletion EMSA, chromatin immunoprecipitation assays, Western blotting, protein translocation, protein phosphorylation measurements. Understanding the molecular basis of HNE induction of GCL to assist development of rational design of a drug with a less potential toxicity is the long range goal.