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Proc Natl Acad Sci U S A. 2008 Apr 8;105(14):5609-13. doi: 10.1073/pnas.0801456105. Epub 2008 Apr 2.

Postendocytotic traffic of the galanin R1 receptor: a lysosomal signal motif on the cytoplasmic terminus.

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1
Departments of Neuroscience and Physiology and Pharmacology, Karolinska Institutet, S-171 77 Stockholm, Sweden.

Abstract

The neuropeptide galanin R1 receptor (GalR1) was tagged at its C terminus with EGFP (GalR1-EGFP) to study receptor localization and trafficking. In PC12 and HEK293 cells, functional GalR1-EGFP was expressed on the plasma membrane and internalized into cytoplasmic vesicles after galanin stimulation. The internalization was blocked by 0.4 M sucrose and by silencing of clathrin with siRNA methodology. Internalized GalR1-EGFP and LysoTracker, a lysosomal marker, overlapped in intracellular vesicles after prolonged galanin stimulation. This colocalization was strongly reduced after site-directed mutagenesis of the motif YXXØ on the C terminus of GalR1 (where Ø is a bulky hydrophobic residue and X any amino acid). Taken together, these data suggest that GalR1 is internalized via the clathrin-dependent, endocytic pathway and then, to a large extent, delivered to lysosomes for degradation through the lysosome-targeting signal YXXØ.

PMID:
18385373
PMCID:
PMC2291124
DOI:
10.1073/pnas.0801456105
[Indexed for MEDLINE]
Free PMC Article
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