Morphology and structure of lipoproteins revealed by an optimized negative-staining protocol of electron microscopy

Lei Zhang, James Song, Giorgio Cavigiolio, Brian Y. Ishida, Shengli Zhang, John P. Kane, Karl H. Weisgraber, Michael N. Oda, Kerry Anne Rye, Henry J. Pownall, Gang Ren

Research output: Contribution to journalArticle

78 Scopus citations

Abstract

Plasma lipoprotein levels are predictors of risk for coronary artery disease. Lipoprotein structure-function relationships provide important clues that help identify the role of lipoproteins in cardiovascular disease. The compositional and conformational heterogeneity of lipoproteins are major barriers to the identification of their structures, as discovered using traditional approaches. Although electron microscopy (EM) is an alternative approach, conventional negative staining (NS) produces rouleau artifacts. In a previous study of apolipoprotein (apo)E4-containing reconstituted HDL (rHDL) particles, we optimized the NS method in a way that eliminated rouleaux. Here we report that phosphotungstic acid at high buffer salt concentrations plays a key role in rouleau formation. We also validate our protocol for analyzing the major plasma lipoprotein classes HDL, LDL, IDL, and VLDL, as well as homogeneously prepared apoA-I-containing rHDL. High-contrast EM images revealed morphology and detailed structures of lipoproteins, especially apoA-I-containing rHDL, that are amenable to three-dimensional reconstruction by single-particle analysis and electron tomography. -Zhang, L., J. Song, G. Cavigiolio, B. Y. Ishida, S. Zhang, J. P. Kane, K. H. Weisgraber, M. N. Oda, K-A. Rye, H. J. Pownall, and G. Ren. The morphology and structure of lipoproteins revealed by an optimized negativestaining protocol of electron microscopy.

Original languageEnglish (US)
Pages (from-to)175-184
Number of pages10
JournalJournal of lipid research
Volume52
Issue number1
DOIs
StatePublished - Jan 2011

Keywords

  • Lipoprotein morphology
  • Lipoprotein structure
  • Protocol

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Endocrinology

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