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J. Biol. Chem., Vol. 266, Issue 13, 8560-8566, 05, 1991
AW Brandli, ED Adamson and K Simons
Madin-Darby canine kidney (MDCK) cells polarize and generate distinct
apical and basolateral membrane domains when grown on permeable filter
supports. Under these conditions, they transcytose fluid-phase markers.
Recently, receptor-mediated transcytosis of epidermal growth factor (EGF)
across MDCK cells has been reported (Maratos-Flier, E., Kao, C.- Y. Y.,
Verdin, E. M., and King, G. L. (1987) J. Cell Biol. 105, 1595- 1601). We
examined the role of the EGF receptor in this process. Transcytosis of EGF
occurred only in the basolateral-to-apical direction, was time-dependent,
and inhibited by the addition of unlabeled EGF in a concentration-dependent
manner. In contrast to previous work, we found that only about 5% of
basolaterally bound EGF was transported to the apical chamber. The
half-time of transport was 90 min. A mutant cell line of MDCK, MDCKII-RCAr,
was used to study the expression of the EGF receptor. Cell surface
glycoproteins of these mutant cells can be efficiently labeled with
[3H]galactose by exogalactosylation. The EGF receptor was found to be
expressed only on the basolateral surface. Addition of EGF to the
basolateral medium resulted in rapid internalization and degradation of the
receptor. Testing directly for transcytosis of basolateral glycoproteins,
we detected several proteins transported across the cell. The EGF receptor,
however, was not among this group of proteins. Taking these results
together, we suggest the following model. Internalization of EGF on the
basolateral surface is mediated by the EGF receptor. EGF dissociates from
the receptor in an endocytic compartment. A fraction of the EGF is then
diverted nonselectively to the transcytotic pathway, as found for other
fluid-phase markers previously (Bomsel, M., Prydz, K., Parton, R. G.,
Gruenberg, J., and Simons, K. (1989) J. Cell Biol. 109, 3243-3258.
Transcytosis of epidermal growth factor. The epidermal growth factor receptor mediates uptake but not transcytosis
European Molecular Biology Laboratory, Cell Biology Programme, Heidelberg, Germany.
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