Biogenetic Pathways of Plasma Membrane Proteins in Caco-2, a Human Intestinal Epithelial Cell Line

• Published in: The Journal of cell biology 1990-10, Vol.111 (4), p.1351-1361
• Main Authors: Le Bivic, André, Quaroni, Andrea, Nichols, Buford, Rodriguez-Boulan, Enrique
• Format: Article
• Language: English
• Subjects: Alkaline Phosphatase - metabolism; Aminopeptidases - metabolism; Analytical, structural and metabolic biochemistry; Antibodies; Antigens; Assessors parcel numbers; Biological and medical sciences; Biological Transport - physiology; Biotin; Caco 2 cells; CD13 Antigens; Cell lines; Cell membranes; Cells; Colonic Neoplasms - metabolism; Epithelial cells; epithelium; Fundamental and applied biological sciences. Psychology; Hepatocytes; Humans; Intestinal Mucosa - metabolism; intestine; Kidney cells; membrane proteins; Membrane Proteins - metabolism; Miscellaneous; Protein Processing, Post-Translational; Proteins; Sucrase-Isomaltase Complex - metabolism; Tumor Cells, Cultured
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.111.4.1351

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 (>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.