Format

Send to

Choose Destination
Biochemistry. 1983 Nov 22;22(24):5667-74.

Prelysosomal divergence of transferrin and epidermal growth factor during receptor-mediated endocytosis.

Abstract

The routes followed by epidermal growth factor and transferrin during their endocytosis by human epithelial cells were compared in double-label studies by using density gradient centrifugation of cell homogenates and fluorescence microscopy with intact cells. Gradient centrifugation studies of cells incubated with radioactively labeled epidermal growth factor and transferrin indicated that both ligands initially were associated with a class of vesicles having a density of 1.037 g/mL and then were rapidly transferred to a membrane compartment having a slightly higher density (1.039 g/mL). Subsequently, the two ligands diverged. Epidermal growth factor ultimately was transferred to a membranous compartment containing lysosomal enzymes (density (1.08 g/mL) where it was degraded. Transferrin was released intact from the cells; very little was transferred to lysosomes. Using fluorescently labeled ligands, it was observed that after cells were warmed to 37 degrees C for 5 min, transferrin and epidermal growth factor gave coincident, punctate fluorescent patterns, strongly suggesting they were localized within the same endocytic vesicles. Subsequently, the epidermal growth factor signal was observed in lysosomes whereas the transferrin signal became weaker and diffuse and did not coincide with the punctate epidermal growth factor fluorescence. The time course of the divergence of the radioactive and fluorescent ligands coupled with the previous morphologic studies on the pathway of epidermal growth factor internalization [Willingham, M. C., & Pastan, I. (1982) J. Cell Biol. 94, 207-212] suggests that the sorting process is prelysosomal and possibly Golgi associated.

PMID:
6317024
DOI:
10.1021/bi00293a033
[Indexed for MEDLINE]

Supplemental Content

Loading ...
Support Center