Lipoprotein assembly. Apolipoprotein B size determines lipoprotein core circumference.
Apolipoprotein B (apoB) is an essential structural protein for the two triglyceride-rich lipoproteins synthesized by humans: chylomicrons and very low density lipoproteins. Although much is known about the role of apoB in clearance of lipoproteins from the circulation, relatively little is known about its role in the assembly of nascent lipoproteins. Therefore, we have investigated the relationship between the length of various N-terminal apoB fragments and the characteristics of the lipoproteins with which these fragments were associated. After the addition of puromycin, HepG2 cells secreted a discrete series of C-terminally truncated apoB fragments on lipoprotein particles including apoB25, apoB29, apoB31, apoB33, apoB36, apoB38, apoB42, apoB45, apoB49, apoB51, apoB55, apoB70, and apoB80. Also, using plasmids encoding apoB26, apoB33, apoB37, apoB42, and apoB48, C-terminally truncated apoB fragments were expressed and secreted after transient transfection of HepG2 cells. Lipoproteins bearing the metabolically labeled apoB fragments were isolated from the cell culture media and characterized in terms of size, density, flotation coefficient, and composition. Lipoprotein radii, calculated from their flotation coefficients and buoyant densities, were used to derive the circumference of the non-polar core of each lipoprotein species. When plotted as a function of apoB size, core circumference defined a straight line of near-zero intercept. The slope of this line was approximately 1 A of core circumference/1 kDa of apoB molecular mass. A model for the mechanism of lipoprotein assembly in HepG2 cells, consistent with the concept that apoB size determines lipoprotein core circumference, is proposed.
Spring
D J
DJ
Department of Chemistry and Biochemistry, University of California, Los Angeles 90024.
Chen-Liu
L W
LW
Chatterton
J E
JE
Elovson
J
J
Schumaker
V N
VN
D001055:Apolipoproteins B / Q000737:chemistry* / Q000235:genetics; D006528:Carcinoma, Hepatocellular / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008049:Lipase / Q000378:metabolism; D008074:Lipoproteins / Q000737:chemistry* / Q000648:ultrastructure; D008099:Liver / Q000201:enzymology; D008113:Liver Neoplasms / Q000378:metabolism; D008854:Microscopy, Electron; D008970:Molecular Weight; D010957:Plasmids; D011691:Puromycin / Q000494:pharmacology; D014162:Transfection; D014407:Tumor Cells, Cultured
D001055:Apolipoproteins B; D008074:Lipoproteins; D011691:Puromycin; D008049:Lipase