Immunofunctional properties of a yolk sac epithelial cell line expressing two proteins gp280 and gp330 of the intermicrovillar area of proximal tubule cells: inhibition of endocytosis by the specific antibodies.
The apical domain of epithelial cells lining the proximal tubule and the yolk sac is characterized by the development of extensive microvilli which limit intermicrovillar spaces backed on their cytoplasmic aspect by a coat of clathrin. These membrane areas which give rise to endocytic vesicles are characterized by the expression on their outer aspect of two high molecular weight glycoproteins: gp330 and gp280. In this study we report on an epithelial cell line, BN/MSV, derived from a yolk sac carcinoma which expresses these two glycoproteins. By indirect immunofluorescence, gp330 and gp280 were detectable on the cell surface and after permeabilization in intracytoplasmic vesicles. At the ultrastructural level they were concentrated in clathrin-coated membrane areas and although gp280 could also be detected in non-coated areas. The two proteins were synthesized independently in the form of high molecular weight polymers by biosynthetically labeled BN/MSV cells. Both were released in the supernatant, but, in spite of previously reported similarities by peptide mapping, only gp330 coprecipitated with a 45 kDa protein comigrating with the alpha 2-macroglobulin receptor-associated protein (MRAP). Culture of the cells in the presence of antibodies to gp280 and to a lesser extent of antibodies to gp330 inhibited the internalization of [14C]sucrose and peroxidase. When followed intracellularly at the ultrastructural level, the compartments containing peroxidase in the presence of anti-gp280 or gp330 antibodies were morphologically distinct from those observed under control conditions: vesicles were of smaller size and irregular shape and accumulation in lysosomes was delayed.(ABSTRACT TRUNCATED AT 250 WORDS)
Le Panse
S
S
INSERM, U064, Hôpital Tenon, Paris France.
Galceran
M
M
Pontillon
F
F
Lelongt
B
B
van de Putte
M
M
Ronco
P M
PM
Verroust
P J
PJ
D000906:Antibodies; D000911:Antibodies, Monoclonal; D002277:Carcinoma; D002462:Cell Membrane / Q000737:chemistry; D003034:Coated Pits, Cell-Membrane / Q000737:chemistry; D003593:Cytoplasm / Q000737:chemistry; D004705:Endocytosis; D011992:Endosomes / Q000201:enzymology; D004847:Epithelial Cells; D026541:Heymann Nephritis Antigenic Complex; D006735:Horseradish Peroxidase / Q000378:metabolism; D007687:Kidney Tubules, Proximal / Q000737:chemistry; D008562:Membrane Glycoproteins / Q000032:analysis / Q000096:biosynthesis / Q000276:immunology / Q000502:physiology*; D008871:Microvilli / Q000737:chemistry; D013395:Sucrose / Q000378:metabolism; D014407:Tumor Cells, Cultured; D015017:Yolk Sac / Q000166:cytology / Q000378:metabolism*
D000906:Antibodies; D000911:Antibodies, Monoclonal; D026541:Heymann Nephritis Antigenic Complex; D008562:Membrane Glycoproteins; D013395:Sucrose; D006735:Horseradish Peroxidase