Juxtamembrane region of the amino terminus of the corticotropin releasing factor receptor type 1 is important for ligand interaction

I. Q. Assil, Jun Qi Lai Jun Qi, M. Arai, M. Shomali, A. B. Abou-Samra

Research output: Contribution to journalArticle

Abstract

The functional properties of the amino terminus (NT) of the corticotropin releasing factor (CRF) receptor type 1 (R1) were studied by use of murine (m) CRFR1 and rat (r) parathyroid hormone (PTH)/parathyroid hormone-related peptide receptor (PTH1R) chimeras. The chimeric receptor CXP, in which the NT of mCRFR1 was annealed to the TMs of PTH1R, and the reciprocal hybrid, PXC, bound radiolabeled analogues of sauvagine and PTH(3-34), respectively. Neither hybrid bound radiolabeled CRF or PTH(1-34). CRF and PTH(1-34) weakly stimulated intracellular cAMP accumulation in COS-7 cells transfected with PXC and CXP, respectively. Thus the NT is required for ligand binding and the TMs are required for agonist-stimulated cAMP accumulation. Replacing individual intercysteine segments of PXC with their mCRFR1 counterparts did not rescue CRF or sauvagine radioligand binding or stimulation of cAMP accumulation. Replacement of residues 1-31 of mCRFR 1 with their PTH 1R counterparts resulted in a chimeric receptor, PEC, which had normal CRFR1 functional properties. In addition, a series of chimeras (F1PEC-F6PEC) were generated by replacement of the NT intercysteine residues of PEC with their PTH1R counterparts. Only F1PEC, F2PEC, and F3PEC showed detectable CRF and sauvagine radioligand binding. All of the PEC chimeras except F5PEC increased cAMP accumulation. These data indicate that the Cys68-Glu109 domain is important for binding and that the Cys87-Cys102 region plays an important role in CRFR1 activation.

Original languageEnglish (US)
Pages (from-to)1187-1195
Number of pages9
JournalBiochemistry
Volume40
Issue number5
DOIs
StatePublished - Feb 6 2001
Externally publishedYes

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Corticotropin-Releasing Hormone
Ligands
Parathyroid Hormone
Parathyroid Hormone Receptor Type 1
Rats
COS Cells
Chemical activation
CRF receptor type 1
sauvagine

ASJC Scopus subject areas

  • Biochemistry

Cite this

Juxtamembrane region of the amino terminus of the corticotropin releasing factor receptor type 1 is important for ligand interaction. / Assil, I. Q.; Lai Jun Qi, Jun Qi; Arai, M.; Shomali, M.; Abou-Samra, A. B.

In: Biochemistry, Vol. 40, No. 5, 06.02.2001, p. 1187-1195.

Research output: Contribution to journalArticle

Assil, I. Q. ; Lai Jun Qi, Jun Qi ; Arai, M. ; Shomali, M. ; Abou-Samra, A. B. / Juxtamembrane region of the amino terminus of the corticotropin releasing factor receptor type 1 is important for ligand interaction. In: Biochemistry. 2001 ; Vol. 40, No. 5. pp. 1187-1195.
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AB - The functional properties of the amino terminus (NT) of the corticotropin releasing factor (CRF) receptor type 1 (R1) were studied by use of murine (m) CRFR1 and rat (r) parathyroid hormone (PTH)/parathyroid hormone-related peptide receptor (PTH1R) chimeras. The chimeric receptor CXP, in which the NT of mCRFR1 was annealed to the TMs of PTH1R, and the reciprocal hybrid, PXC, bound radiolabeled analogues of sauvagine and PTH(3-34), respectively. Neither hybrid bound radiolabeled CRF or PTH(1-34). CRF and PTH(1-34) weakly stimulated intracellular cAMP accumulation in COS-7 cells transfected with PXC and CXP, respectively. Thus the NT is required for ligand binding and the TMs are required for agonist-stimulated cAMP accumulation. Replacing individual intercysteine segments of PXC with their mCRFR1 counterparts did not rescue CRF or sauvagine radioligand binding or stimulation of cAMP accumulation. Replacement of residues 1-31 of mCRFR 1 with their PTH 1R counterparts resulted in a chimeric receptor, PEC, which had normal CRFR1 functional properties. In addition, a series of chimeras (F1PEC-F6PEC) were generated by replacement of the NT intercysteine residues of PEC with their PTH1R counterparts. Only F1PEC, F2PEC, and F3PEC showed detectable CRF and sauvagine radioligand binding. All of the PEC chimeras except F5PEC increased cAMP accumulation. These data indicate that the Cys68-Glu109 domain is important for binding and that the Cys87-Cys102 region plays an important role in CRFR1 activation.

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