Journal
JOURNAL OF LIPID RESEARCH
Volume 52, Issue 1, Pages 175-184Publisher
ELSEVIER
DOI: 10.1194/jlr.D010959
Keywords
lipoprotein structure; lipoprotein morphology; protocol
Categories
Funding
- Office of Science, Office of Basic Energy Sciences of the U.S. Department of Energy [DE-AC02-05CH11231]
- W.M. Keck foundations
- State Scholarship of China through China Scholarship Council [2008628018]
- NATIONAL HEART, LUNG, AND BLOOD INSTITUTE [R01HL115153] Funding Source: NIH RePORTER
- NATIONAL INSTITUTE OF GENERAL MEDICAL SCIENCES [R01GM104427] Funding Source: NIH RePORTER
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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. J. Lipid Res. 2011. 52: 175-184.
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