Endocytosis of receptor-bound insulin-like growth factor II is enhanced by mannose-6-phosphate in IM9 cells.
The insulin-like growth factor II (IGF-II), and glycoproteins containing mannose 6-phosphate (M6P), bind to two different sites of the same receptor molecule (Morgan et al, Nature 329:301, 1987). To study the interactions between the two ligands on their common receptor in intact cells, we examined the effect of free M6P on IGF-II binding and endocytosis in the IM9 human lymphoblastoid cell line. M6P, up to a 3 mM concentration, had no effect on the binding of IGF-II to the cell surface receptor of intact IM9 cells at 4 degrees C. By contrast, when IM9 cells were incubated with 125I-IGF-II at 37 degrees C, 1mM M6P increased cell-associated radioactivity by twofold. The increase was resistant to acid wash at 4 degrees C, and therefore assumed to represent endocytosed IGF-II. Acid-washable radioactivity was no different, confirming that, in intact cells, M6P does not affect IGF-II surface binding. In addition, preincubation of cells with M6P at 37 degrees C for up to 3 hours did not change the abundance of receptor on the cell surface, as measured by a subsequent 4 degrees C binding assay. We conclude that M6P causes a shift of IGF-II-occupied receptors form the cell surface to intracellular locations without affecting surface binding of this ligand in IM9 cells. The effect could be produced by the binding of M6P itself, or by the displacement of endogenous phosphomannosylated ligands.
Polychronakos
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Department of Pediatrics, McGill University, Montreal, Quebec, Canada.
Piscina
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D002460:Cell Line; D002738:Chloroquine / Q000494:pharmacology; D004705:Endocytosis / Q000187:drug effects*; D006600:Hexosephosphates / Q000494:pharmacology*; D006801:Humans; D007335:Insulin-Like Growth Factor II / Q000378:metabolism*; D007700:Kinetics; D008360:Mannosephosphates / Q000494:pharmacology*; D011972:Receptor, Insulin / Q000378:metabolism*; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000378:metabolism*
D006600:Hexosephosphates; D008360:Mannosephosphates; D017451:Receptors, Somatomedin; D013002:Somatomedins; C027693:mannose-6-phosphate; D007335:Insulin-Like Growth Factor II; D002738:Chloroquine; D011972:Receptor, Insulin