Biogenetic pathways of plasma membrane proteins in Caco-2, a human intestinal epithelial cell line.
We studied the sorting and surface delivery of three apical and three basolateral proteins in the polarized epithelial cell line Caco-2, using pulse-chase radiolabeling and surface domain-selective biotinylation (Le Bivic, A., F. X. Real, and E. Rodriguez-Boulan. 1989. Proc. Natl. Acad. Sci. USA. 86:9313-9317). While the basolateral proteins (antigen 525, HLA-I, and transferrin receptor) were targeted directly and efficiently to the basolateral membrane, the apical markers (sucrase-isomaltase [SI], aminopeptidase N [APN], and alkaline phosphatase [ALP]) reached the apical membrane by different routes. The large majority (80%) of newly synthesized ALP was directly targeted to the apical surface and the missorted basolateral pool was very inefficiently transcytosed. SI was more efficiently targeted to the apical membrane (greater than 90%) but, in contrast to ALP, the missorted basolateral pool was rapidly transcytosed. Surprisingly, a distinct peak of APN was detected on the basolateral domain before its accumulation in the apical membrane; this transient basolateral pool (at least 60-70% of the enzyme reaching the apical surface, as measured by continuous basal addition of antibodies) was efficiently transcytosed. In contrast with their transient basolateral expression, apical proteins were more stably localized on the apical surface, apparently because of their low endocytic capability in this membrane. Thus, compared with two other well-characterized epithelial models, MDCK cells and the hepatocyte, Caco-2 cells have an intermediate sorting phenotype, with apical proteins using both direct and indirect pathways, and basolateral proteins using only direct pathways, during biogenesis.
Le Bivic
A
A
Department of Cell Biology and Anatomy, Cornell University Medical College, New York 10021.
Quaroni
A
A
Nichols
B
B
Rodriguez-Boulan
E
E
D000469:Alkaline Phosphatase / Q000378:metabolism; D000626:Aminopeptidases / Q000378:metabolism; D001692:Biological Transport / Q000502:physiology; D001710:Biotin; D018826:CD13 Antigens; D003110:Colonic Neoplasms / Q000378:metabolism; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D013394:Sucrase-Isomaltase Complex / Q000378:metabolism; D014407:Tumor Cells, Cultured
D008565:Membrane Proteins; D001710:Biotin; D000469:Alkaline Phosphatase; D013394:Sucrase-Isomaltase Complex; D000626:Aminopeptidases; D018826:CD13 Antigens