Apical secretion of apolipoproteins from enterocytes.
Synthesis and secretion of apolipoproteins in pig small intestine was studied by pulse-chase labeling of jejunal segments, kept in organ culture. Apo A-1 and apo B-48 were the two major proteins released, constituting 25 and 10%, respectively, of the total amount of labeled protein in the mucosal-side medium where they appeared with a t1/2 of 50-60 min. Using tissue from fasting animals, > 85% of newly synthesized apo A-1 and about one third of apo B-48 was released to the mucosal-side medium. Newly synthesized apolipoprotein that remained associated with the intestinal segment accumulated in the soluble fraction, suggesting a basolateral secretion into the intercellular space, and both this accumulation and the release to the medium was prevented by culture at 20 degrees C. The specific radioactivity of apo A-1 and apo B-48 released to the medium was significantly higher than that of the corresponding apolipoproteins remaining associated with the intestinal tissue. Furthermore, during culture periods of up to 5 h, the enterocytes and their tight junctions largely remained intact as evidenced by the inaccessibility of the nonpermeable surface marker Ruthenium red. We therefore propose that enterocytes release most of their newly made free apo A-1 and a significant portion of apo B-48 by exocytosis via the brush border membrane into the intestinal lumen. Fat absorption reduced apolipoprotein secretion to the medium and induced the formation of chylomicrons, containing apo A-1 at their surface, as evidenced by immunogold electron microscopy. The chylomicrons were localized in the Golgi complex and near the basolateral plasma membrane, but not in the apical region of the enterocytes, indicating that only free apolipoproteins are secreted to the intestinal lumen.
Danielsen
E M
EM
Department of Biochemistry, Panum Institute, University of Copenhagen, Denmark.
Hansen
G H
GH
Poulsen
M D
MD
D000818:Animals; D001053:Apolipoproteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D003600:Cytosol / Q000378:metabolism / Q000648:ultrastructure; D004847:Epithelial Cells; D004848:Epithelium / Q000378:metabolism / Q000648:ultrastructure; D005110:Extracellular Space / Q000378:metabolism; D007365:Intercellular Junctions / Q000648:ultrastructure; D007413:Intestinal Mucosa / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D007421:Intestine, Small / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D007700:Kinetics; D008715:Methionine / Q000378:metabolism; D008854:Microscopy, Electron; D008871:Microvilli / Q000378:metabolism / Q000648:ultrastructure; D008970:Molecular Weight; D009924:Organ Culture Techniques; D013552:Swine; D013997:Time Factors
D001053:Apolipoproteins; D008715:Methionine