Barwell, James, Conner, Alex C. and Poyner, David R.. (2011) Extracellular loops 1 and 3 and their associated transmembrane regions of the calcitonin receptor-like receptor are needed for CGRP receptor function. Biochimica et Biophysica Acta - Molecular Cell Research, Vol.1813 (No.10). pp. 1906-1916. ISSN 0167-4889

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Abstract

The first and third extracellular loops (ECL) of G protein-coupled receptors (GPCRs) have been implicated in ligand binding and receptor function. This study describes the results of an alanine/leucine scan of ECLs 1 and 3 and loop-associated transmembrane (TM) domains of the secretin-like GPCR calcitonin receptor-like receptor which associates with receptor activity modifying protein 1 to form the CGRP receptor. Leu195Ala, Val198Ala and Ala199Leu at the top of TM2 all reduced alpha CGRP-mediated cAMP production and internalization: Leu195Ala and Ala199Leu also reduced alpha CGRP binding. These residues form a hydrophobic cluster within an area defined as the "minor groove" of rhodopsin-like GPCRs. Within ECL1, Ala203Leu and Ala206Leu influenced the ability of alpha CGRP to stimulate adenylate cyclase. In TM3, His219Ala, Leu220Ala and Leu222Ala have influences on alpha CGRP binding and cAMP production; they are likely to indirectly influence the binding site for alpha CGRP as well as having an involvement in signal transduction. On the exofacial surfaces of TMs 6 and 7, a number of residues were identified that reduced cell surface receptor expression, most noticeably Leu351Ala and Glu357Ala in TM6. The residues may contribute to the RAMP1 binding interface. Ile360Ala impaired alpha CGRP-mediated cAMP production. Ile360 is predicted to be located close to ECL2 and may facilitate receptor activation. Identification of several crucial functional loci gives further insight into the activation mechanism of this complex receptor system and may aid rational drug design.

Item Type:	Journal Article
Subjects:	Q Science > QP Physiology Q Science > QR Microbiology
Divisions:	Faculty of Medicine > Warwick Medical School
Library of Congress Subject Headings (LCSH):	G proteins -- Receptors, Radioligand assay, Ligand binding (Biochemistry), Cellular signal transduction
Journal or Publication Title:	Biochimica et Biophysica Acta - Molecular Cell Research
Publisher:	Elsevier Science BV
ISSN:	0167-4889
Date:	October 2011
Volume:	Vol.1813
Number:	No.10
Number of Pages:	11
Page Range:	pp. 1906-1916
Identification Number:	10.1016/j.bbamcr.2011.06.005
Status:	Peer Reviewed
Publication Status:	Published
Access rights to Published version:	Restricted or Subscription Access
Funder:	British Heart Foundation
Grant number:	FS/05/054 (BHF)
References:	[1] J. Barwell, P.S. Miller, D. Donnelly, D.R. Poyner, Mapping interaction sites within the N-terminus of the calcitonin gene-related peptide receptor; the role of residues 23– 60 of the calcitonin receptor-like receptor, Peptides 31 (2010) 170–176. [2] C. Bergwitz, S.A. Jusseaume, M.D. Luck, H. Juppner, T.J. Gardella, Residues in the membrane-spanning and extracellular loop regions of the parathyroid hormone (PTH)-2 receptor determine signaling selectivity for PTH and PTH-related peptide, J. Biol. Chem. 272 (1997) 28861–28868. [3] A.C. Conner, D.L. Hay, J. Simms, S.G. Howitt,M. Schindler,D.M. Smith, et al., A key role for transmembrane prolines in calcitonin receptor-like receptor agonist binding and signalling: implications for family B G-protein-coupled receptors, Mol. Pharmacol. 67 (2005) 20–31. [4] A.C. Conner, J. Simms, S.G. Howitt, M. Wheatley, D.R. Poyner, Multiple residues within the second extracellular loop of the CL receptor are required for receptor activation by CGRP pA2 Online, Vol. 3, 2005, p. 99. [5] M. Conner, M.R. Hicks, T. Dafforn, T.J. Knowles, C. Ludwig, S. Staddon, et al., Functional and biophysical analysis of the C-terminus of the CGRP-receptor; a family B GPCR, Biochemistry 47 (2008) 8434–8444. [6] E.Di Paolo, J.P.Vilardaga,H. Petry,N.Moguilevsky,A. Bollen, P. Robberecht, et al., Role of charged amino acids conserved in the vasoactive intestinal polypeptide/secretin family of receptors on the secretin receptor functionality, Peptides 20 (1999) 1187–1193. [7] T.J. Dolinsky, P. Czodrowski, H. Li, J.E. Nielsen, J.H. Jensen, G. Klebe, et al., PDB2PQR: expanding and upgrading automated preparation of biomolecular structures for molecular simulations, Nucleic Acids Res. 35 (2007) W522–W525. [8] M. Dong, R.F. Cox, L.J. Miller, Juxtamembranous region of the amino terminus of the family B G protein-coupled calcitonin receptor plays a critical role in small-molecule agonist action, J. Biol. Chem. 284 (2009) 21839–21847. [9] M. Dong, P.C. Lam, D.I. Pinon, A. Orry, P.M. Sexton, R. Abagyan, et al., Secretin occupies a single protomer of the homodimeric secretin receptor complex: insights from photoaffinity labeling studies using dual sites of covalent attachment, J. Biol. Chem. 285 (2010) 9919–9931. [10] K. Du, P. Nicole, A. Couvineau, M. Laburthe, Aspartate 196 in the first extracellular loop of the human VIP1 receptor is essential for VIP binding and VIP-stimulated cAMP production, Biochem. Biophys. Res. Commun. 230 (1997) 289–292. [11] K.G. Harikumar, J. Simms, G. Christopoulos, P.M. Sexton, L.J.Miller, Molecular basis of association of receptor activity-modifying protein 3 with the family B G proteincoupled secretin receptor, Biochemistry 48 (2009) 11773–11785. [12] S. Hilairet, C. Belanger, J. Bertrand, A. Laperriere, S.M. Foord, M. Bouvier, Agonistpromoted internalization of a ternary complex between calcitonin receptor-like receptor, receptor activity-modifying protein 1 (RAMP1), and beta-arrestin, J. Biol. Chem. 276 (2001) 42182–42190. [13] M.H. Holtmann, S. Ganguli, E.M. Hadac, V.Dolu, L.J. Miller,Multiple extracellular loop domains contribute critical determinants for agonist binding and activation of the secretin receptor, J. Biol. Chem. 271 (1996) 14944–14949. [14] W. Im, M. Feig, C.L. Brooks 111, An implicit membrane generalized born theory for the study of structure, stability, and interactions of membrane proteins, Biophys. J. 85 (2003) 2900–2918. [15] I. Langer, P. Vertongen, J. Perret, M. Waelbroeck, P. Robberecht, Lysine 195 and aspartate 196 in the first extracellular loop of the VPAC1 receptor are essential for high affinity binding of agonists but not of antagonists, Neuropharmacology 44 (2003) 125–131. [16] R.A. Laskowski, PDBsum: summaries and analyses of PDB structures, Nucleic Acids Res. 29 (2001) 221–222. [17] R.A. Laskowski, D.S. Moss, J.M. Thornton, Main-chain bond lengths and bond angles in protein structures, J. Mol. Biol. 231 (1993) 1049–1067. [18] H. Li, A.D. Robertson, J.H. Jensen, Very fast empirical prediction and rationalization of protein pKa values, Proteins 61 (2005) 704–721. [19] L.M. McLatchie, N.J. Fraser, M.J. Main, A.Wise, J. Brown, N. Thompson, et al., RAMPs regulate the transport and ligand specificity of the calcitonin-receptor-like receptor, Nature 393 (1998) 333–339. [20] L.J. Miller, Q. Chen, P.C. Lam, D.I. Pinon, P.M. Sexton, R. Abagyan, et al., Refinement of glucagon-like peptide 1 docking to its intact receptor using mid-region photolabile probes and molecular modeling, J. Biol. Chem. 286 (2011) 15895–15907. [21] P. Monaghan, B.E. Thomas, I. Woznica, A. Wittelsberger, D.F. Mierke, M. Rosenblatt, Mapping peptide hormone–receptor interactions using a disulfide-trapping approach, Biochemistry 47 (2008) 5889–5895. [22] C. Parthier, S. Reedtz-Runge, R. Rudolph, M.T. Stubbs, Passing the baton in class B GPCRs: peptide hormone activation via helix induction? Trends Biochem. Sci. 34 (2009) 303–310. [23] M.C. Peeters, G.J. van Westen, Q. Li, A.P. Ijzerman, Importance of the extracellular loops in G protein-coupled receptors for ligand recognition and receptor activation, Trends Pharmacol. Sci. 32 (2011) 35–42. [24] M.M. Rosenkilde, T. Benned-Jensen, T.M. Frimurer, T.W. Schwartz, The minor binding pocket: amajor player in 7TMreceptor activation, Trends Pharmacol. Sci. 31 (2010) 567–574. [25] S. Runge, C. Gram, H. Brauner-Osborne, K. Madsen, L.B. Knudsen, B.S. Wulff, Three distinct epitopes on the extracellular face of the glucagon receptor determine specificity for the glucagon amino terminus, J. Biol. Chem. 278 (2003) 28005–28010. [26] S.P. Sheikh, J.P. Vilardarga, T.J. Baranski, O. Lichtarge, T. Iiri, E.C. Meng, et al., Similar structures and shared switch mechanisms of the beta2-adrenoceptor and the parathyroid hormone receptor. Zn(II) bridges between helices III and VI block activation, J. Biol. Chem. 274 (1999) 17033–17041. [27] J. Simms, D.L. Hay, R.J. Bailey, G. Konycheva, G. Bailey, M. Wheatley, et al., Structure–function analysis of RAMP1 by alanine mutagenesis, Biochemistry 48 (2009) 198–205. [28] C.S. Soto, M. Fasnacht, J. Zhu, L. Forrest, B. Honig, Loop modeling: sampling, filtering, and scoring, Proteins 70 (2008) 834–843. [29] E. ter Haar, C.M. Koth, N. Abdul-Manan, L. Swenson, J.T. Coll, J.A. Lippke, et al., Crystal structure of the ectodomain complex of the CGRP receptor, a class-B GPCR, reveals the site of drug antagonism, Structure 18 (2010) 1083–1093. [30] S. Vohra, S.V. Chintapalli, C.J. Illingworth, P.J. Reeves, P.M. Mullineaux, H.S. Clark, et al., Computational studies of family A and family B GPCRs, Biochem. Soc. Trans. 35 (2007) 749–754. [31] Z. Xiang, C.S. Soto, B. Honig, Evaluating conformational free energies: the colony energy and its application to the problem of loop prediction, Proc. Natl. Acad. Sci. U. S. A. 99 (2002) 7432–7437. [32] F. Xu, H. Wu, V. Katritch, G.W. Han, K.A. Jacobson, Z.G. Gao, et al., Structure of an agonist-bound human A2A adenosine receptor, Science 332 (2011) 322–327.
URI:	http://wrap.warwick.ac.uk/id/eprint/38738

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