Receptors coupled to pertussis toxin-sensitive G-proteins traffic to opposite surfaces in Madin-Darby canine kidney cells A1 adenosine receptors achieve apical and α2A adrenergic receptors achieve basolateral localization

Christine Saunders, Jeffrey R. Keefer, Amy P. Kennedy, Jack N. Wells, Lee E. Limbird

Research output: Contribution to journalArticlepeer-review

Abstract

The α2A adrenergic receptor (α2AAR) previously was shown to be directly delivered to and retained on the lateral subdomain of renal epithelial cells. The present studies demonstrate that, in contrast, wild-type and epitope-tagged canine A1 adenosine receptors (A1AdoR) are apically enriched (65-83%) in Madin-Darby canine kidney (MDCKII) and porcine renal epithelial (LLC-PKI) cells, based on surface biotinylation strategies detecting photoaffinity-labeled A1AdoR. Confocal microscopy corroborated the apical enrichment of the epitopetagged A1AdoR. Metabolic labeling studies revealed that this steady-state polarization is achieved by direct delivery to both the apical (60-75%) and basolateral surface. Growth of A1AdoR-expressing cells as monolayers was achieved only following Transwell culture in the presence of A1AdoR antagonists, which decreased cell growth, suggesting that A1AdoR elicit MDCKII cell proliferation. The preferential apical but detectable basolateral localization of A1AdoR provides a molecular understanding of published reports that functional responses can be elicited following apical as well as basolateral delivery of adenosine agonists in varying renal preparations. These findings also suggest that receptor chimeras derived from the Gi/Go-protein-coupled α2AAR and A1AdoR will be informative in revealing structural features critical for basolateral versus apical targeting.

Original languageEnglish (US)
Pages (from-to)995-1002
Number of pages8
JournalJournal of Biological Chemistry
Volume271
Issue number2
DOIs
StatePublished - Jan 12 1996

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

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