Proteomic analysis of human macrophages exposed to hypochlorite-oxidized low-density lipoprotein

Biochim Biophys Acta. 2009 Mar;1794(3):446-58. doi: 10.1016/j.bbapap.2008.11.015. Epub 2008 Dec 6.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Blotting, Western
  • Cells, Cultured
  • Electrophoresis, Gel, Two-Dimensional
  • Foam Cells / drug effects
  • Foam Cells / metabolism*
  • Humans
  • Hypochlorous Acid / pharmacology*
  • Lipoproteins, LDL / chemistry*
  • Macrophages / drug effects
  • Macrophages / metabolism
  • Proteomics*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Lipoproteins, LDL
  • oxidized low density lipoprotein
  • Hypochlorous Acid