Home LiteratureArticle Details
PMID: 19022181 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

Ligand-binding architecture of human CB2 cannabinoid receptor: evidence for receptor subtype-specific binding motif and modeling GPCR activation.

Chemistry & biology ·Vol. 15 ·No. 11 ·2008-11-24 ·Pages 1207-19

Pei Y, Mercier RW, Anday JK, Thakur GA, Zvonok AM, Hurst D, Reggio PH, Janero DR, Makriyannis A

Abstract

The extensive physiological influence of transmission through the CB2 cannabinoid receptor makes this G protein-coupled receptor (GPCR) a promising therapeutic target for treating neuropathic pain, inflammation, and immune disorders. However, there is little direct structural information pertaining to either GPCR or CB2-receptor ligand recognition and activation. The present work helps characterize experimentally the ligand-binding interactions of the human CB2 (hCB2) receptor. This study illustrates how our overall experimental approach, "ligand-assisted protein structure" (LAPS), affords direct determination of the requirements for ligand binding to the hCB2 receptor and discrimination among the binding motifs for ligands that activate therapeutically relevant GPCRs.

MeSH Terms
Affinity Labels/metabolism Amino Acid Sequence Cell Line Drug Discovery Gene Expression Regulation Humans Ligands Models, Molecular Molecular Sequence Data Mutagenesis, Site-Directed Mutant Proteins/chemistry,genetics,metabolism Mutation Protein Binding Receptor, Cannabinoid, CB1/metabolism Receptor, Cannabinoid, CB2/chemistry,genetics,metabolism Substrate Specificity
Chemicals
Affinity Labels Ligands Mutant Proteins Receptor, Cannabinoid, CB1 Receptor, Cannabinoid, CB2
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Pei Ying
School of Pharmacy, University of Connecticut, Storrs, CT 06269, USA.
Mercier Richard W
Anday Jenine K
Thakur Ganesh A
Zvonok Alexander M
Hurst Dow
Reggio Patricia H
Janero David R
Makriyannis Alexandros
References (59)
59 references, click to expand
  1. A novel electrophilic high affinity irreversible probe for the cannabinoid receptor.
    Life Sci. 1995;56(23-24):1957-62 PMID: 7776819
  2. Pharmacotherapeutic targeting of the endocannabinoid signaling system: drugs for obesity and the metabolic syndrome.
    Physiol Behav. 2008 Mar 18;93(4-5):671-86 PMID: 18155257
  3. Mutational analysis and molecular modelling of the antagonist SR 144528 binding site on the human cannabinoid CB(2) receptor.
    Eur J Pharmacol. 2000 Jul 28;401(1):17-25 PMID: 10915832
  4. Novel, not adenylyl cyclase-coupled cannabinoid binding site in cerebellum of mice.
    Biochem Biophys Res Commun. 2002 Mar 22;292(1):231-5 PMID: 11890697
  5. CB1 cannabinoid antagonists: structure-activity relationships and potential therapeutic applications.
    Curr Top Med Chem. 2008;8(3):205-30 PMID: 18289089
  6. Covalent inhibitors of human monoacylglycerol lipase: ligand-assisted characterization of the catalytic site by mass spectrometry and mutational analysis.
    Chem Biol. 2008 Aug 25;15(8):854-62 PMID: 18721756
  7. F200A substitution in the third transmembrane helix of human cannabinoid CB1 receptor converts AM2233 from receptor agonist to inverse agonist.
    Eur J Pharmacol. 2006 Feb 15;531(1-3):41-6 PMID: 16438957
  8. Identification of the binding sites of the SR49059 nonpeptide antagonist into the V1a vasopressin receptor using sulfydryl-reactive ligands and cysteine mutants as chemical sensors.
    J Biol Chem. 2003 Oct 10;278(41):40010-9 PMID: 12869559
  9. Beta2 adrenergic receptor activation. Modulation of the proline kink in transmembrane 6 by a rotamer toggle switch.
    J Biol Chem. 2002 Oct 25;277(43):40989-96 PMID: 12167654
  10. Crystal structure of the ligand-free G-protein-coupled receptor opsin.
    Nature. 2008 Jul 10;454(7201):183-7 PMID: 18563085
  11. N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxamide (SR141716A) interaction with LYS 3.28(192) is crucial for its inverse agonism at the cannabinoid CB1 receptor.
    Mol Pharmacol. 2002 Dec;62(6):1274-87 PMID: 12435794
  12. Comparative protein modelling by satisfaction of spatial restraints.
    J Mol Biol. 1993 Dec 5;234(3):779-815 PMID: 8254673
  13. A ternary complex model explains the agonist-specific binding properties of the adenylate cyclase-coupled beta-adrenergic receptor.
    J Biol Chem. 1980 Aug 10;255(15):7108-17 PMID: 6248546
  14. The conformation, location, and dynamic properties of the endocannabinoid ligand anandamide in a membrane bilayer.
    J Biol Chem. 2005 Aug 19;280(33):29788-95 PMID: 15964843
  15. Constitutive activation of the beta2 adrenergic receptor alters the orientation of its sixth membrane-spanning segment.
    J Biol Chem. 1997 Jul 25;272(30):18546-9 PMID: 9228019
  16. Role of a conserved lysine residue in the peripheral cannabinoid receptor (CB2): evidence for subtype specificity.
    Mol Pharmacol. 1999 Mar;55(3):605-13 PMID: 10051546
  17. Cannabinergic ligands.
    Chem Phys Lipids. 2002 Dec 31;121(1-2):3-19 PMID: 12505686
  18. (-)-7'-Isothiocyanato-11-hydroxy-1',1'-dimethylheptylhexahydrocannabinol (AM841), a high-affinity electrophilic ligand, interacts covalently with a cysteine in helix six and activates the CB1 cannabinoid receptor.
    Mol Pharmacol. 2005 Dec;68(6):1623-35 PMID: 16157695
  19. Recent advances in the development of selective ligands for the cannabinoid CB(2) receptor.
    Curr Top Med Chem. 2008;8(3):187-204 PMID: 18289088
  20. Small angle X-ray diffraction and differential scanning calorimetric studies on O-methyl-(-)-delta 8-tetrahydrocannabinol and its 5' iodinated derivative in membrane bilayers.
    Biochim Biophys Acta. 1995 Jul 26;1237(2):183-8 PMID: 7632712
  21. Cysteine 2.59(89) in the second transmembrane domain of human CB2 receptor is accessible within the ligand binding crevice: evidence for possible CB2 deviation from a rhodopsin template.
    Mol Pharmacol. 2005 Jul;68(1):69-83 PMID: 15840841
  22. The two-state model of receptor activation.
    Trends Pharmacol Sci. 1995 Mar;16(3):89-97 PMID: 7540781
  23. Mutation of a highly conserved aspartate residue in the second transmembrane domain of the cannabinoid receptors, CB1 and CB2, disrupts G-protein coupling.
    J Pharmacol Exp Ther. 1998 May;285(2):651-8 PMID: 9580609
  24. Crystal structure of rhodopsin: A G protein-coupled receptor.
    Science. 2000 Aug 4;289(5480):739-45 PMID: 10926528
  25. A lysine residue of the cannabinoid receptor is critical for receptor recognition by several agonists but not WIN55212-2.
    Mol Pharmacol. 1996 May;49(5):891-6 PMID: 8622639
  26. Chemical rescue of phosphoryl transfer in a cavity mutant: a cautionary tale for site-directed mutagenesis.
    Biochemistry. 2001 Jan 16;40(2):403-13 PMID: 11148034
  27. Computational methods in drug design: modeling G protein-coupled receptor monomers, dimers, and oligomers.
    AAPS J. 2006 May 12;8(2):E322-36 PMID: 16796383
  28. Cannabinoids activate an inwardly rectifying potassium conductance and inhibit Q-type calcium currents in AtT20 cells transfected with rat brain cannabinoid receptor.
    J Neurosci. 1995 Oct;15(10):6552-61 PMID: 7472417
  29. Molecular characterization of a peripheral receptor for cannabinoids.
    Nature. 1993 Sep 2;365(6441):61-5 PMID: 7689702
  30. Milk intake and survival in newborn cannabinoid CB1 receptor knockout mice: evidence for a "CB3" receptor.
    Eur J Pharmacol. 2003 Feb 7;461(1):27-34 PMID: 12568912
  31. High-resolution crystal structure of an engineered human beta2-adrenergic G protein-coupled receptor.
    Science. 2007 Nov 23;318(5854):1258-65 PMID: 17962520
  32. Agonist-induced conformational changes at the cytoplasmic side of transmembrane segment 6 in the beta 2 adrenergic receptor mapped by site-selective fluorescent labeling.
    J Biol Chem. 2001 Mar 23;276(12):9279-90 PMID: 11118431
  33. Enzyme catalysis: removing chemically 'essential' residues by site-directed mutagenesis.
    Trends Biochem Sci. 2001 Aug;26(8):497-503 PMID: 11504626
  34. Potent cannabinergic indole analogues as radioiodinatable brain imaging agents for the CB1 cannabinoid receptor.
    J Med Chem. 2005 Oct 6;48(20):6386-92 PMID: 16190764
  35. Cannabinoid CB1 receptor of cat cerebral arterial muscle functions to inhibit L-type Ca2+ channel current.
    Am J Physiol. 1999 Jun;276(6):H2085-93 PMID: 10362691
  36. Cannabinoid receptor activation differentially regulates the various adenylyl cyclase isozymes.
    J Neurochem. 1998 Oct;71(4):1525-34 PMID: 9751186
  37. Rat brain cannabinoid receptor modulates N-type Ca2+ channels in a neuronal expression system.
    Mol Pharmacol. 1996 Apr;49(4):707-14 PMID: 8609900
  38. Targeting the cannabinoid CB2 receptor: modelling and structural determinants of CB2 selective ligands.
    Br J Pharmacol. 2008 Jan;153(2):335-46 PMID: 17982473
  39. Evaluation of the in vivo receptor occupancy for the behavioral effects of cannabinoids using a radiolabeled cannabinoid receptor agonist, R-[125/131I]AM2233.
    Synapse. 2006 Aug;60(2):93-101 PMID: 16715483
  40. Structural features of the central cannabinoid CB1 receptor involved in the binding of the specific CB1 antagonist SR 141716A.
    J Biol Chem. 1996 Mar 22;271(12):6941-6 PMID: 8636122
  41. 5'-Azido-delta 8-THC: a novel photoaffinity label for the cannabinoid receptor.
    J Med Chem. 1992 Aug 7;35(16):3076-9 PMID: 1323683
  42. CB2 cannabinoid receptor agonists: pain relief without psychoactive effects?
    Curr Opin Pharmacol. 2003 Feb;3(1):62-7 PMID: 12550743
  43. Requirement of rigid-body motion of transmembrane helices for light activation of rhodopsin.
    Science. 1996 Nov 1;274(5288):768-70 PMID: 8864113
  44. Structure-activity relationships of pyrazole derivatives as cannabinoid receptor antagonists.
    J Med Chem. 1999 Feb 25;42(4):769-76 PMID: 10052983
  45. A mutation-induced activated state of the beta 2-adrenergic receptor. Extending the ternary complex model.
    J Biol Chem. 1993 Mar 5;268(7):4625-36 PMID: 8095262
  46. SR144528 as inverse agonist of CB2 cannabinoid receptor.
    J Vet Sci. 2002 Sep;3(3):179-84 PMID: 12514329
  47. Small angle x-ray diffraction studies on the topography of cannabinoids in synaptic plasma membranes.
    Pharmacol Biochem Behav. 1991 Nov;40(3):547-52 PMID: 1666918
  48. Structure of a beta1-adrenergic G-protein-coupled receptor.
    Nature. 2008 Jul 24;454(7203):486-91 PMID: 18594507
  49. (-)-11-Hydroxy-7'-isothiocyanato-1',1'-dimethylheptyl-delta 8-THC: a novel, high-affinity irreversible probe for the cannabinoid receptor in the brain.
    J Med Chem. 1994 Nov 11;37(23):3867-70 PMID: 7966145
  50. Ligand binding and modulation of cyclic AMP levels depend on the chemical nature of residue 192 of the human cannabinoid receptor 1.
    J Neurochem. 1998 Jan;70(1):366-73 PMID: 9422383
  51. Targeting the cannabinoid CB2 receptor: mutations, modeling and development of CB2 selective ligands.
    Curr Med Chem. 2005;12(10):1217-37 PMID: 15892633
  52. Signaling pathway associated with stimulation of CB2 peripheral cannabinoid receptor. Involvement of both mitogen-activated protein kinase and induction of Krox-24 expression.
    Eur J Biochem. 1996 May 1;237(3):704-11 PMID: 8647116
  53. Characterization of a region of steric interference at the cannabinoid receptor using the active analog approach.
    J Med Chem. 1993 Jun 11;36(12):1761-71 PMID: 8510104
  54. Purification and mass spectroscopic analysis of human CB1 cannabinoid receptor functionally expressed using the baculovirus system.
    J Pept Res. 2005 Sep;66(3):138-50 PMID: 16083441
  55. Structural diversity of G protein-coupled receptors and significance for drug discovery.
    Nat Rev Drug Discov. 2008 Apr;7(4):339-57 PMID: 18382464
  56. Ligand based structural studies of the CB1 cannabinoid receptor.
    J Pept Res. 2002 Dec;60(6):348-56 PMID: 12464113
  57. Functional role of serine residues of transmembrane dopamin VII in signal transduction of CB2 cannabinoid receptor.
    J Vet Sci. 2002 Sep;3(3):185-91 PMID: 12514330
  58. Construction of a 3D model of the cannabinoid CB1 receptor: determination of helix ends and helix orientation.
    Life Sci. 1995;56(23-24):1971-82 PMID: 7776821
  59. Specific tryptophan UV-absorbance changes are probes of the transition of rhodopsin to its active state.
    Biochemistry. 1996 Aug 27;35(34):11149-59 PMID: 8780519
Article Info
Journal
Chemistry & biology
Abbr.
Chem Biol
ISSN
1879-1301
DOI
10.1016/j.chembiol.2008.10.011
Published
2008-11-24
Pages
1207-19
Language
English
Region
United States
NLM ID
9500160
PMCID
PMC3700404
Subset
IM
Grants
NIDA NIH HHS · R01 DA003934 · United States
NIDA NIH HHS · R37 DA003801 · United States
NIDA NIH HHS · P01 DA009158 · United States
NIDA NIH HHS · DA9158 · United States
NIDA NIH HHS · R01 DA003801 · United States
NIDA NIH HHS · DA03934 · United States
Intramural NIH HHS · Z01 DA000489 · United States
NIDA NIH HHS · K02 DA000489 · United States
NIDA NIH HHS · DA3801 · United States
External Links
PubMed source DOI: 10.1016/j.chembiol.2008.10.011 PMC Full Text
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: product@genelibs.com

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2