Optimized negative-staining electron microscopy for lipoprotein studies

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Optimized negative-staining electron microscopy for lipoprotein studies.

BACKGROUND Negative-staining (NS), a rapid, simple and conventional technique of electron microscopy (EM), has been commonly used to initially study the morphology and structure of proteins for half a century. Certain NS protocols however can cause artifacts, especially for structurally flexible or lipid-related proteins, such as lipoproteins. Lipoproteins were often observed in the form of rou...

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Apolipoprotein E (apoE), one of the major protein components of lipoproteins in the peripheral and central nervous systems, regulates cholesterol metabolism through its interaction with members of the low density lipoprotein receptor family. One key to understanding apoE function is determining the structure of lipid-bound forms of apoE. Negative-staining (NS) electron microscopy (EM) is an eas...

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

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. Althou...

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Section Staining for Electron Microscopy

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Structural determination of proteins is rather challenging for proteins with molecular masses between 40 - 200 kDa. Considering that more than half of natural proteins have a molecular mass between 40 - 200 kDa, a robust and high-throughput method with a nanometer resolution capability is needed. Negative staining (NS) electron microscopy (EM) is an easy, rapid, and qualitative approach which h...

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ژورنال

عنوان ژورنال: Biochimica et Biophysica Acta (BBA) - General Subjects

سال: 2013

ISSN: 0304-4165

DOI: 10.1016/j.bbagen.2012.09.016