title: Proteomic analysis of human macrophages exposed to hypochlorite-oxidized low-density lipoprotein.
abstract: The invasion of monocytes through the endothelial wall of arteries and their transformation from macrophage into form cells has been implicated as a critical initiating event in atherogenesis. Human THP-1 monocytic cells can be induced to differentiate into macrophages by phorbol myristate acetate (PMA) treatment, and can be converted into foam cells by exposure to oxidized low-density lipoprotein (oxLDL). To identify proteins potentially involved in atherosclerotic processes, we performed a proteomic analysis of THP-1 macrophages exposed to oxLDL generated by treatment with native LDL with hypochlorous acid/hypochlorite (HOCl/OCl(-)). We detected more than a thousand proteins, of which 104 differentially expressed proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF) and the NCBI database. The largest differences in expression were observed for bifunctional purine biosynthesis protein, vacuolar protein sorting 33A, breast carcinoma amplified sequence, adenine phosphoribosyltransferase, and tropomyosin alpha 3 chain. Interestingly, many apoptotic proteins such as lamin B1, poly (ADP-ribose) polymerase, Bcl-2 related protein A1 and vimentin were identified by MALDI-TOF analysis. Identities were confirmed by matching the sequence of several tryptic peptides using MALDI-TOF/TOF MS, Western blot analyses and immunofluorescent microscopy. The data described here will contribute to establishing a functional profile of the human macrophage proteome. Furthermore, the proteins identified in this study are attractive candidates for further biomarkers involved in the pathogenesis of atherosclerosis.
authors.affiliation: Department of Life Sciences and Biotechnology, School of Life Sciences, Kyungpook National University, Daegu, 702-701, Republic of Korea.
authors.forename: Jeong Han
authors.initials: JH
authors.lastname: Kang
authors.forename: Hyun Su
authors.initials: HS
authors.lastname: Ryu
authors.forename: Hyun Tae
authors.initials: HT
authors.lastname: Kim
authors.forename: Su Jin
authors.initials: SJ
authors.lastname: Lee
authors.forename: Ung-Kyu
authors.initials: UK
authors.lastname: Choi
authors.forename: Yong Bok
authors.initials: YB
authors.lastname: Park
authors.forename: Tae-Lin
authors.initials: TL
authors.lastname: Huh
authors.forename: Myung-Sook
authors.initials: MS
authors.lastname: Choi
authors.forename: Tae-Cheon
authors.initials: TC
authors.lastname: Kang
authors.forename: Soo Young
authors.initials: SY
authors.lastname: Choi
authors.forename: Oh-Shin
authors.initials: OS
authors.lastname: Kwon
mesh_terms: D015153:Blotting, Western; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D005487:Foam Cells / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology*; D008077:Lipoproteins, LDL / Q000737:chemistry*; D008264:Macrophages / Q000187:drug effects / Q000378:metabolism; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D006997:Hypochlorous Acid