Proteomic analysis of human macrophages exposed to hypochlorite-oxidized low-density lipoprotein.
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.
Kang
Jeong Han
JH
Department of Life Sciences and Biotechnology, School of Life Sciences, Kyungpook National University, Daegu, 702-701, Republic of Korea.
Ryu
Hyun Su
HS
Kim
Hyun Tae
HT
Lee
Su Jin
SJ
Choi
Ung-Kyu
UK
Park
Yong Bok
YB
Huh
Tae-Lin
TL
Choi
Myung-Sook
MS
Kang
Tae-Cheon
TC
Choi
Soo Young
SY
Kwon
Oh-Shin
OS
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
D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D006997:Hypochlorous Acid