Basolateral sorting signal of the 300-kDa mannose 6-phosphate receptor.
In polarized cells, the delivery of numerous membrane proteins from the trans-Golgi network to the basolateral surface depends on specific sequences located in their cytoplasmic domain. We have previously shown that the insulin-like growth factor-II/mannose 6-phosphate receptor (IGF-II/MPR) exhibits a polarized cell surface distribution in the human colon adenocarcinoma (Caco-2) cell line in which there is a threefold enrichment on the basolateral surface. To investigate the role of residues in the cytoplasmic region of the receptor that facilitates its entry into the basolateral sorting pathway, we generated stably transfected Caco-2 cell lines expressing various mutant bovine IGF-II/MPRs. The steady-state surface distribution of mutant receptors was analyzed by subjecting filter-grown cell monolayers to incubation with iodinated IGF-II/MPR-specific antibody or to indirect immunofluorescence and visualization by confocal microscopy. Together, these results demonstrate that the sorting of the IGF-II/MPR to the basolateral cell surface depends on recognition of sequences located in its cytoplasmic region that are distinct from the Tyr-based internalization and dileucine-dependent endosomal trafficking motifs.
Wick
Debra A
DA
Department of Biochemistry, Medical College of Wisconsin, Milwaukee, Wisconsin 53226, USA.
Seetharam
Bellur
B
Dahms
Nancy M
NM
D000595:Amino Acid Sequence; D000818:Animals; D018938:Caco-2 Cells; D002417:Cattle; D016764:Cell Polarity / Q000502:physiology; D015870:Gene Expression; D006801:Humans; D007413:Intestinal Mucosa / Q000166:cytology; D007930:Leucine / Q000235:genetics; D016296:Mutagenesis / Q000502:physiology; D021382:Protein Sorting Signals / Q000502:physiology*; D017434:Protein Structure, Tertiary; D017527:Receptor, IGF Type 2 / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D014162:Transfection
D021382:Protein Sorting Signals; D017527:Receptor, IGF Type 2; D007930:Leucine