Disulfide bonds are required for folding and secretion of apolipoprotein B regardless of its lipidation state.
Apolipoprotein (apo) B-100, an essential protein for the assembly and secretion of very low density lipoproteins depends on lipid binding (lipidation) for its secretion. Seven of its 8 disulfides are clustered within the N-terminal 21%. The role of these disulfides in the secretion of lipidated or unlipidated truncated forms of apoB was studied in C127 cells expressing apoB-17, apoB-29, or apoB-41. These cells do not express microsomal triglyceride transfer protein yet secrete apoB-41 on triacylglycerol-rich lipoproteins while apoB-29 and apoB-17 are secreted with little or no lipid, respectively. Dithiothreitol utilized in pulse-chase studies prevented the cotranslational formation of disulfides and when added posttranslationally reduced native disulfides. As a result, the secretion of reduced apoB forms was blocked and they were retained in the cells. Reduced apoB polypeptides were rescued following removal of dithiothreitol, as they underwent post-translational disulfide bonding, attained their mature form, and were subsequently secreted. Together the data suggest that in C127 cells the formation of native disulfides is critical for the folding and secretion of apoB independent of its length, its requirement for lipidation or microsomal triglyceride transfer protein expression. Therefore, these cells provide an appropriate model to study the folding of apoB in great detail.
Burch
W L
WL
Department of Biophysics, Center for Advanced Biomedical Research, Boston University School of Medicine, Boston, Massachusetts 02118, USA.
Herscovitz
H
H
D000818:Animals; D001055:Apolipoproteins B / Q000737:chemistry* / Q000378:metabolism; D004220:Disulfides / Q000737:chemistry*; D004229:Dithiothreitol / Q000494:pharmacology; D008055:Lipids / Q000737:chemistry*; D051379:Mice; D014176:Protein Biosynthesis; D017510:Protein Folding; D014407:Tumor Cells, Cultured
D001055:Apolipoproteins B; D004220:Disulfides; D008055:Lipids; D004229:Dithiothreitol