Proteomic characterization of human plasma high density lipoprotein fractionated by gel filtration chromatography

Scott M. Gordon, Jingyuan Deng, L. Jason Lu, W. Sean Davidson

Research output: Contribution to journalArticlepeer-review

188 Scopus citations

Abstract

Plasma levels of high density lipoprotein cholesterol (HDL-C) are inversely proportional to the incidence of cardiovascular disease. Recent applications of modern proteomic technologies have identified upward of 50 distinct proteins associated with HDL particles with many of these newly discovered proteins implicating HDL in nonlipid transport processes including complement activation, acute phase response and innate immunity. However, almost all MS-based proteomic studies on HDL to date have utilized density gradient ultracentrifugation techniques for HDL isolation prior to analysis. These involve high shear forces and salt concentrations that can disrupt HDL protein interactions and alter particle function. Here, we used high-resolution size exclusion chromatography to fractionate normal human plasma to 17 phospholipid-containing subfractions. Then, using a phospholipid binding resin, we identified proteins that associate with lipoproteins of various sizes by electrospray ionization mass spectrometry. We identified 14 new phospholipid-associated proteins that migrate with traditionally defined HDL, several of which further support roles for HDL in complement regulation and protease inhibition. The increased fractionation inherent to this method allowed us to visualize HDL protein distribution across particle size with unprecedented resolution. The observed heterogeneity across subfractions suggests the presence of HDL particle subpopulations each with distinct protein components that may prove to impart distinct physiological functions.

Original languageEnglish
Pages (from-to)5239-5249
Number of pages11
JournalJournal of Proteome Research
Volume9
Issue number10
DOIs
StatePublished - Oct 1 2010

Keywords

  • apolipoprotein
  • high density lipoprotein
  • lipoprotein
  • mass spectrometry
  • proteomics

ASJC Scopus subject areas

  • Biochemistry
  • General Chemistry

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