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Coupling of the PTH/PTHrP receptor to multiple G-proteins

Direct demonstration of receptor activation of Gs′ Gq/11′ and Gi(1) by [α-32P]GTP-γ-Azidoanilide photoaffinity labeling

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Abstract

Parathyroid hormone (PTH) elicits many of its physiological effects by activating distinct G-proteincoupled signaling cascades that lead to synthesis of cyclic AMP and hydrolysis of phosphatidylinositol 4,5-bisphosphate. Using the nonhydrolyzable photoreactive GTP analog [α-32P]GTP-γ-azidoanilide (GTP-AA) and peptide antisera raised against G-protein α-subunits, we studied coupling of the PTH receptor to G-proteins in rat osteoblast-like cells (ROS 17/2.8), and in human embryonal kidney cells expressing the cloned human PTH/parathyroid hormone-related peptide (PTHrP) receptor at 40,000 receptors/cell (C20) or 400,000 receptors/cell (C21). Incubation of C21 membranes (but not C20 membranes) with [Nle8,18, Tyr34]-bovine PTH(1-34) amide (bPTH[1-34]) led to concentration-dependent incorporation of GTP-AA into the two isoforms of Gα s, into Gα q/11, and to a much lesser extent into Gα i(1). In ROS 17/2.8 cells, bPTH(1-34) increased the incorporation of GTP-AA into Gα s, but not into Gα q/11 or Gα i. The ability of bPTH(1-34) to increase labeling of Gα s and Gα q/11 was correlated with the receptor-dependent sensitivity of the adenylyl cyclase and phospholipase C signaling pathways to the hormone.

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Authors and Affiliations

  1. Division of Rheumatology, Johns Hopkins University School of Medicine, Baltimore, MD

    Helen Robinson & Joan M. Bathon

  2. Division of Bone and Mineral Metabolism, Beth Israel Hospital, Boston, MA

    Mark Pines & Larry J. Suva

  3. Division of Endocrinology and Metabolism, Johns Hopkins University School of Medicine, Ross 1029, 720 Rutland Ave., 21205, Baltimore, MD

    William F. Schwindinger, Janine Fredericks, Lakrisha Watkins & Michael A. Levine

Authors
  1. William F. Schwindinger
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  2. Janine Fredericks
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  3. Lakrisha Watkins
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  4. Helen Robinson
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  5. Joan M. Bathon
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  6. Mark Pines
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  7. Larry J. Suva
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Correspondence to William F. Schwindinger.

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Schwindinger, W.F., Fredericks, J., Watkins, L. et al. Coupling of the PTH/PTHrP receptor to multiple G-proteins. Endocr 8, 201–209 (1998). https://doi.org/10.1385/ENDO:8:2:201

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  • DOI: https://doi.org/10.1385/ENDO:8:2:201

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