Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

Pair your accounts.

Export articles to Mendeley

Get article recommendations from ACS based on references in your Mendeley library.

You’ve supercharged your research process with ACS and Mendeley!

Click to create an ACS ID

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

Your Mendeley pairing has expired. Please reconnect
ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Juxtamembrane Region of the Amino Terminus of the Corticotropin Releasing Factor Receptor Type 1 Is Important for Ligand Interaction

View Author Information
Endocrine Unit, Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts 02114
Cite this: Biochemistry 2001, 40, 5, 1187–1195
Publication Date (Web):January 10, 2001
Copyright © 2001 American Chemical Society

    Article Views





    Other access options


    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 mCRFR1 with their PTH1R 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.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.


     Corresponding author:  Telephone (617) 726-6723; fax (617) 726-1703; e-mail [email protected].

    Cited By

    This article is cited by 32 publications.

    1. Oliver Kraetke,, Brian Holeran,, Hartmut Berger,, Emanuel Escher,, Michael Bienert, and, Michael Beyermann. Photoaffinity Cross-Linking of the Corticotropin-Releasing Factor Receptor Type 1 with Photoreactive Urocortin Analogues. Biochemistry 2005, 44 (47) , 15569-15577.
    2. Sam R. J. Hoare,, Sue K. Sullivan,, David A. Schwarz,, Nicholas Ling,, Wylie W. Vale,, Paul D. Crowe, and, Dimitri E. Grigoriadis. Ligand Affinity for Amino-Terminal and Juxtamembrane Domains of the Corticotropin Releasing Factor Type I Receptor:  Regulation by G-Protein and Nonpeptide Antagonists. Biochemistry 2004, 43 (13) , 3996-4011.
    3. Kaushik Sarkar, Lisa Joedicke, Marta Westwood, Rebecca Burnley, Michael Wright, David McMillan, Bernadette Byrne. Modulation of PTH1R signaling by an extracellular binding antibody. 2022, 109-132.
    4. Jan M. Deussing, Alon Chen. The Corticotropin-Releasing Factor Family: Physiology of the Stress Response. Physiological Reviews 2018, 98 (4) , 2225-2286.
    5. D.E. Grigoriadis. Corticotropin-Releasing Factor and Urocortin Receptors. 2017, 57-65.
    6. Syed Jalal Khundmiri, Rebecca D. Murray, Eleanor Lederer. PTH and Vitamin D. 2016, 561-601.
    7. Dimitris K Grammatopoulos. Insights into mechanisms of corticotropin‐releasing hormone receptor signal transduction. British Journal of Pharmacology 2012, 166 (1) , 85-97.
    8. Irene Coin, Marilyn H. Perrin, Wylie W. Vale, Lei Wang. Photo‐Cross‐Linkers Incorporated into G‐Protein‐Coupled Receptors in Mammalian Cells: A Ligand Comparison. Angewandte Chemie International Edition 2011, 50 (35) , 8077-8081.
    9. Irene Coin, Marilyn H. Perrin, Wylie W. Vale, Lei Wang. Photo‐Cross‐Linkers Incorporated into G‐Protein‐Coupled Receptors in Mammalian Cells: A Ligand Comparison. Angewandte Chemie 2011, 123 (35) , 8227-8231.
    10. . Fundamental Principles of Cell‐Based Assays. 2010, 213-238.
    11. Kostas Gkountelias, Theodoros Tselios, Maria Venihaki, George Deraos, Iakovos Lazaridis, Olga Rassouli, Achille Gravanis, George Liapakis. Alanine Scanning Mutagenesis of the Second Extracellular Loop of Type 1 Corticotropin-Releasing Factor Receptor Revealed Residues Critical for Peptide Binding. Molecular Pharmacology 2009, 75 (4) , 793-800.
    12. Iman Assil-Kishawi, Tareq A. Samra, Dale F. Mierke, Abdul B. Abou-Samra. Residue 17 of Sauvagine Cross-links to the First Transmembrane Domain of Corticotropin-releasing Factor Receptor 1 (CRFR1). Journal of Biological Chemistry 2008, 283 (51) , 35644-35651.
    13. Malin C. Lagerström, Helgi B. Schiöth. Structural diversity of G protein-coupled receptors and significance for drug discovery. Nature Reviews Drug Discovery 2008, 7 (4) , 339-357.
    14. Angela Wittelsberger, Michael Rosenblatt. Parathyroid Hormone–Receptor Interactions. 2008, 595-637.
    15. Dimitri E. Grigoriadis, Samuel R. J. Hoare. Emerging Anxiolytics: Corticotropin‐Releasing Factor Receptor Antagonists. 2007
    16. E. L. Vodovozova. Photoaffinity labeling and its application in structural biology. Biochemistry (Moscow) 2007, 72 (1) , 1-20.
    17. H Berger, N Heinrich, D Wietfeld, M Bienert, M Beyermann. Evidence that corticotropin‐releasing factor receptor type 1 couples to Gs‐ and Gi‐proteins through different conformations of its J‐domain. British Journal of Pharmacology 2006, 149 (7) , 942-947.
    18. Edward W. Hillhouse, Dimitris K. Grammatopoulos. The Molecular Mechanisms Underlying the Regulation of the Biological Activity of Corticotropin-Releasing Hormone Receptors: Implications for Physiology and Pathophysiology. Endocrine Reviews 2006, 27 (3) , 260-286.
    19. Christy R. R. Grace, Marilyn H. Perrin, Michael R. DiGruccio, Charleen L. Miller, Jean E. Rivier, Wylie W. Vale, Roland Riek. NMR structure and peptide hormone binding site of the first extracellular domain of a type B1 G protein-coupled receptor. Proceedings of the National Academy of Sciences 2004, 101 (35) , 12836-12841.
    20. Frank M. Dautzenberg, Sandra Wille. Binding differences of human and amphibian corticotropin-releasing factor type 1 (CRF1) receptors: identification of amino acids mediating high-affinity astressin binding and functional antagonism. Regulatory Peptides 2004, 118 (3) , 165-173.
    21. M. Venihaki, S. Sakihara, S. Subramanian, P. Dikkes, S. C. Weninger, G. Liapakis, T. Graf, J. A. Majzoub. Urocortin III, A Brain Neuropeptide of the Corticotropin‐Releasing Hormone Family: Modulation by Stress and Attenuation of Some Anxiety‐Like Behaviours. Journal of Neuroendocrinology 2004, 16 (5) , 411-422.
    22. Vi Pham, Patrick M. Sexton. Photoaffinity scanning in the mapping of the peptide receptor interface of class II G protein—coupled receptors. Journal of Peptide Science 2004, 10 (4) , 179-203.
    23. Tracy L. Bale, Wylie W. Vale. CRF and CRF Receptors: Role in Stress Responsivity and Other Behaviors. Annual Review of Pharmacology and Toxicology 2004, 44 (1) , 525-557.
    24. Marilyn H. Perrin, Michael R. DiGruccio, Steven C. Koerber, Jean E. Rivier, Koichi S. Kunitake, Deborah L. Bain, Wolfgang H. Fischer, Wylie W. Vale. A Soluble Form of the First Extracellular Domain of Mouse Type 2β Corticotropin-releasing Factor Receptor Reveals Differential Ligand Specificity. Journal of Biological Chemistry 2003, 278 (18) , 15595-15600.
    25. Sam R. J. Hoare, Sue K. Sullivan, Nicholas Ling, Paul D. Crowe, Dimitri E. Grigoriadis. Mechanism of Corticotropin-Releasing Factor Type I Receptor Regulation by Nonpeptide Antagonists. Molecular Pharmacology 2003, 63 (3) , 751-765.
    26. Iman Assil-Kishawi, Abdul B. Abou-Samra. Sauvagine Cross-links to the Second Extracellular Loop of the Corticotropin-releasing Factor Type 1 Receptor. Journal of Biological Chemistry 2002, 277 (36) , 32558-32561.
    27. Masako Shimada, Xin Chen, Tomas Cvrk, Helene Hilfiker, Maria Parfenova, Gino V. Segre. Purification and Characterization of a Receptor for Human Parathyroid Hormone and Parathyroid Hormone-related Peptide. Journal of Biological Chemistry 2002, 277 (35) , 31774-31780.
    28. Frank M. Dautzenberg, Jacqueline Higelin, Olaf Brauns, Brigitte Butscha, Richard L. Hauger. Five Amino Acids of the Xenopus laevis CRF (Corticotropin-Releasing Factor) Type 2 Receptor Mediate Differential Binding of CRF Ligands in Comparison with Its Human Counterpart. Molecular Pharmacology 2002, 61 (5) , 1132-1139.
    29. Michael Chorev, Michael Rosenblatt. Parathyroid Hormone-Receptor Interactions. 2002, 423-XXVIII.
    30. K. Eckart, O. Jahn, J. Radulovic, M. Radulovic, T. Blank, O. Stiedl, O. Brauns, H. Tezval, T. Zeyda, J. Spiess. Pharmacology and Biology of Corticotropin-Releasing Factor (CRF) Receptors. Receptors and Channels 2002, 8 (3-4) , 163-177.
    31. M. Chorev. Parathyroid Hormone 1 Receptor: Insights into Structure and Function. Receptors and Channels 2002, 8 (3-4) , 219-242.
    32. John Saunders, John P. Williams. Chapter 3. New developments in the study of corticotropin releasing factor. 2001, 21-30.