Cotranslocational insertion of apolipoprotein B into the inner leaflet of the endoplasmic reticulum.
Apolipoprotein (apo) B100 is required for the distribution of hepatic triglyceride to peripheral tissues as very-low-density lipoproteins. The translocation of apo B100 into the endoplasmic reticulum (ER) and its subsequent assembly into lipoprotein particles is of particular interest as the protein is both very large (relative molecular mass 512,000) and insoluble in water. It has been proposed that apo B translocation occurs in discrete stages and is completed post-translationally. Several sites of arrest of translocation were reported to be present in apo B15 (the N-terminal 15% of the protein). We have re-examined this question by in vitro translation coupled with translocation into microsomes, and find no evidence for transmembrane segments in truncated apo B proteins. Translocated apo B17 is strongly associated with the membrane of the ER, being only partially releasable with alkaline carbonate, and remaining bound to the microsomes following disruption with saponin. The efficient binding of short segments of apo B, despite the absence of transmembrane domains, suggests that apo B is cotranslationally inserted into the inner leaflet of the ER. This will obviate problems caused by the size and insolubility of apo B100, because the growing hydrophobic protein chains will never exist in a lipid-free form during translocation. From the inner leaflet, apo B in association with membrane-derived lipid can bud into the lumen of the ER to form nascent lipoprotein particles.
Pease
R J
RJ
Division of Molecular Medicine, MRC Clinical Research Centre, Harrow, UK.
Harrison
G B
GB
Scott
J
J
D000595:Amino Acid Sequence; D001055:Apolipoproteins B / Q000378:metabolism*; D001483:Base Sequence; D001692:Biological Transport; D004721:Endoplasmic Reticulum / Q000378:metabolism*; D066298:In Vitro Techniques; D008565:Membrane Proteins / Q000378:metabolism*; D008861:Microsomes / Q000378:metabolism; D008969:Molecular Sequence Data; D014176:Protein Biosynthesis; D011499:Protein Processing, Post-Translational; D014158:Transcription, Genetic
D001055:Apolipoproteins B; D008565:Membrane Proteins