Conformation of apolipoprotein B after lipid extraction of low density lipoproteins attached to an electron microscope grid.
We have examined the shape of apolipoprotein B (apoB) from low density lipoproteins (LDL) using a new method to prepare the electron microscope grids. After adsorption of the lipoproteins to a carbon-coated copper grid, lipids were extracted with ethanol-ether 4:1; an aqueous negative stain was then applied. When the LDL residue was examined after this treatment, apoB, together with residual lipid, appeared as an elongated flexible structure about 600-700 A in length consisting of multiple domains of variable width from 20-70 A. Occasionally, the elongated apoB formed an irregular ring-shaped structure, but most of the rings were open. When LDL were pretreated with glutaraldehyde, then adsorbed, extracted, and stained, most of the images were closed rings with an average contour length of 700 A, again consisting of multiple domains of variable sizes. These results are consistent with apoB being composed of multiple domains arranged in an elongated structure on the surface of the LDL, and with distant domains possessing a mutual affinity that favors their cross-linking.
Phillips
M L
ML
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
Schumaker
V N
VN
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D003432:Cross-Linking Reagents / Q000494:pharmacology; D005976:Glutaral / Q000494:pharmacology; D006801:Humans; D008077:Lipoproteins, LDL / Q000032:analysis*; D008854:Microscopy, Electron; D011487:Protein Conformation; D011863:Radioimmunoassay
D000911:Antibodies, Monoclonal; D001055:Apolipoproteins B; D003432:Cross-Linking Reagents; D008077:Lipoproteins, LDL; D005976:Glutaral