Site-specific protein adducts of 4-hydroxy-2(E)-nonenal in human THP-1 monocytic cells: Protein carbonylation is diminished by ascorbic acid
作者： Juan Chavez , Woon-Gye Chung , Cristobal L. Miranda , Mudita Singhal , Jan F. Stevens
DOI: 10.1021/TX9002462
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摘要: The protein targets and sites of modification by 4-hydroxy-2(E)-nonenal (HNE) in human monocytic THP-1 cells after exogenous exposure to HNE were examined using a multipronged proteomic approach involving electrophoretic, immunoblotting, mass spectrometric methods. Immunoblot analysis monoclonal anti-HNE antibodies showed several proteins as adduction. Pretreatment with ascorbic acid resulted reduced levels HNE−protein adducts. Biotinylation Michael-type adducts an aldehyde-reactive hydroxylamine-functionalized probe (aldehyde-reactive probe, ARP) subsequent enrichment facilitated the identification site-specific assignment modifications LC-MS/MS analysis. Sixteen unequivocally identified adduction, eighteen at Cys His residues assigned. involved cytoskeleton organization regulation, p...
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