Identification of a domain in the carboxy terminus of CCK receptor that affects its intracellular trafficking

Y. G O William, Eileen L. Holicky, Elizabeth M. Hadac, Rammohan V. Rao, Laurence J Miller

Research output: Contribution to journalArticle

Abstract

The carboxy-terminal region of many guanine nucleotide-binding protein (G protein)-coupled receptors contains important regulatory sequences such as an NP(x)2-3Y motif, a site of fatty acid acylation, and serine- and threonine-rich domains. The type A CCK receptor contains all of these, yet their significance has not been examined. We have, therefore, constructed a series of receptor site mutants and truncations that interfere with each of these motifs and expressed each in Chinese hamster ovary cells where they were studied for radioligand binding, cell signaling, receptor internalization, and intracellular trafficking. Each construct was synthesized and transported appropriately to the cell surface, where CCK bound with high affinity, elicited an inositol 1,4,5-trisphosphate response, and resulted in internalization and normal trafficking. Thus modification or elimination of each of these established sequence motifs had no substantial effect on any of these parameters of receptor and cellular function. However, an additional construct that truncated the carboxy terminus, eliminating an additional 15-amino-acid segment devoid of any currently recognized sequence motifs, resulted in a marked change in receptor trafficking, with all other parameters of receptor function normal. This mutant receptor construct was delayed at the stage of early endosomes, delaying its progress to the lysosome-enriched perinuclear compartment from the rapid time course followed by wildtype receptor and all of the other constructs. It is proposed that this region of the CCK receptor tail contains a new motif important for intracellular receptor trafficking.

Original languageEnglish (US)
JournalAmerican Journal of Physiology
Volume275
Issue number1 Part1
StatePublished - 1998

Keywords

  • Cholecystokinin
  • Endocytosis
  • G protein-coupled receptor
  • Receptor internalization
  • Receptor trafficking

ASJC Scopus subject areas

  • Physiology (medical)

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