Molecular Determinants for the High Constitutive Activity of the Human Histamine H4 Receptor: Functional Studies on Orthologues and Mutants

Overview

Journal Br J Pharmacol

Publisher Wiley

Specialty Pharmacology

Date 2014 Jun 7

PMID 24903527

Citations 14

Authors

D Wifling

K Loffel

U Nordemann

A Strasser

G Bernhardt

S Dove

R Seifert

A Buschauer

Affiliations

Soon will be listed here.

Abstract

Background And Purpose: Some histamine H4 receptor ligands act as inverse agonists at the human H4 receptor (hH4 R), a receptor with exceptionally high constitutive activity, but as neutral antagonists or partial agonists at the constitutively inactive mouse H4 receptor (mH4 R) and rat H4 receptor (rH4 R). To study molecular determinants of constitutive activity, H4 receptor reciprocal mutants were constructed: single mutants: hH4 R-F169V, mH4 R-V171F, hH4 R-S179A, hH4 R-S179M; double mutants: hH4 R-F169V+S179A, hH4 R-F169V+S179M and mH4 R-V171F+M181S.

Experimental Approach: Site-directed mutagenesis with pVL1392 plasmids containing hH4 or mH4 receptors were performed. Wild-type or mutant receptors were co-expressed with Gαi2 and Gβ1 γ2 in Sf9 cells. Membranes were studied in saturation and competition binding assays ([(3) H]-histamine), and in functional [(35) S]-GTPγS assays with inverse, partial and full agonists of the hH4 receptor.

Key Results: Constitutive activity decreased from the hH4 receptor via the hH4 R-F169V mutant to the hH4 R-F169V+S179A and hH4 R-F169V+S179M double mutants. F169 alone or in concert with S179 plays a major role in stabilizing a ligand-free active state of the hH4 receptor. Partial inverse hH4 receptor agonists like JNJ7777120 behaved as neutral antagonists or partial agonists at species orthologues with lower or no constitutive activity. Some partial and full hH4 receptor agonists showed decreased maximal effects and potencies at hH4 R-F169V and double mutants. However, the mutation of S179 in the hH4 receptor to M as in mH4 receptor or A as in rH4 receptor did not significantly reduce constitutive activity.

Conclusions And Implications: F169 and S179 are key amino acids for the high constitutive activity of hH4 receptors and may also be of relevance for other constitutively active GPCRs.

Linked Articles: This article is part of a themed issue on Histamine Pharmacology Update published in volume 170 issue 1. To view the other articles in this issue visit http://onlinelibrary.wiley.com/doi/10.1111/bph.2013.170.issue-1/issuetoc.

Citing Articles

Structural insights into the agonists binding and receptor selectivity of human histamine H receptor.

Im D, Kishikawa J, Shiimura Y, Hisano H, Ito A, Fujita-Fujiharu Y Nat Commun. 2023; 14(1):6538.

PMID: 37863901 PMC: 10589313. DOI: 10.1038/s41467-023-42260-z.

Label-Free Investigations on the G Protein Dependent Signaling Pathways of Histamine Receptors.

Seibel-Ehlert U, Plank N, Inoue A, Bernhardt G, Strasser A Int J Mol Sci. 2021; 22(18).

PMID: 34575903 PMC: 8467282. DOI: 10.3390/ijms22189739.

The Function of the Histamine H4 Receptor in Inflammatory and Inflammation-Associated Diseases of the Gut.

Schirmer B, Neumann D Int J Mol Sci. 2021; 22(11).

PMID: 34204101 PMC: 8200986. DOI: 10.3390/ijms22116116.

Quantitative Determination and Imaging of Gα Signaling in Live Cells via Split-Luciferase Complementation.

Littmann T, Ozawa T, Bernhardt G Methods Mol Biol. 2021; 2274:69-78.

PMID: 34050463 DOI: 10.1007/978-1-0716-1258-3_7.

Molecular Modeling of Histamine Receptors-Recent Advances in Drug Discovery.

Mehta P, Miszta P, Filipek S Molecules. 2021; 26(6).

PMID: 33810008 PMC: 8004658. DOI: 10.3390/molecules26061778.

References

Nguyen T, Shapiro D, George S, Setola V, Lee D, Cheng R . Discovery of a novel member of the histamine receptor family. Mol Pharmacol. 2001; 59(3):427-33. DOI: 10.1124/mol.59.3.427. View

Reher T, Neumann D, Buschauer A, Seifert R . Incomplete activation of human eosinophils via the histamine H4-receptor: evidence for ligand-specific receptor conformations. Biochem Pharmacol. 2012; 84(2):192-203. DOI: 10.1016/j.bcp.2012.04.004. View

Igel P, Geyer R, Strasser A, Dove S, Seifert R, Buschauer A . Synthesis and structure-activity relationships of cyanoguanidine-type and structurally related histamine H4 receptor agonists. J Med Chem. 2009; 52(20):6297-313. DOI: 10.1021/jm900526h. View

Morse K, Behan J, Laz T, West Jr R, Greenfeder S, Anthes J . Cloning and characterization of a novel human histamine receptor. J Pharmacol Exp Ther. 2001; 296(3):1058-66. View

Leurs R, Chazot P, Shenton F, Lim H, de Esch I . Molecular and biochemical pharmacology of the histamine H4 receptor. Br J Pharmacol. 2009; 157(1):14-23. PMC: 2697796. DOI: 10.1111/j.1476-5381.2009.00250.x. View

Geyer R, Buschauer A . Synthesis and histamine H(3) and H(4) receptor activity of conformationally restricted cyanoguanidines related to UR-PI376. Arch Pharm (Weinheim). 2011; 344(12):775-85. DOI: 10.1002/ardp.201100144. View

Thurmond R, Gelfand E, Dunford P . The role of histamine H1 and H4 receptors in allergic inflammation: the search for new antihistamines. Nat Rev Drug Discov. 2008; 7(1):41-53. DOI: 10.1038/nrd2465. View

Liu C, Wilson S, Kuei C, Lovenberg T . Comparison of human, mouse, rat, and guinea pig histamine H4 receptors reveals substantial pharmacological species variation. J Pharmacol Exp Ther. 2001; 299(1):121-30. View

Cheng Y, Prusoff W . Relationship between the inhibition constant (K1) and the concentration of inhibitor which causes 50 per cent inhibition (I50) of an enzymatic reaction. Biochem Pharmacol. 1973; 22(23):3099-108. DOI: 10.1016/0006-2952(73)90196-2. View

10.

Schneider E, Schnell D, Strasser A, Dove S, Seifert R . Impact of the DRY motif and the missing "ionic lock" on constitutive activity and G-protein coupling of the human histamine H4 receptor. J Pharmacol Exp Ther. 2010; 333(2):382-92. DOI: 10.1124/jpet.109.163220. View

11.

Jablonowski J, Grice C, Chai W, Dvorak C, Venable J, Kwok A . The first potent and selective non-imidazole human histamine H4 receptor antagonists. J Med Chem. 2003; 46(19):3957-60. DOI: 10.1021/jm0341047. View

12.

Nakamura T, Itadani H, Hidaka Y, Ohta M, Tanaka K . Molecular cloning and characterization of a new human histamine receptor, HH4R. Biochem Biophys Res Commun. 2000; 279(2):615-20. DOI: 10.1006/bbrc.2000.4008. View

13.

Rosenbaum D, Zhang C, Lyons J, Holl R, Aragao D, Arlow D . Structure and function of an irreversible agonist-β(2) adrenoceptor complex. Nature. 2011; 469(7329):236-40. PMC: 3074335. DOI: 10.1038/nature09665. View

14.

Shimamura T, Shiroishi M, Weyand S, Tsujimoto H, Winter G, Katritch V . Structure of the human histamine H1 receptor complex with doxepin. Nature. 2011; 475(7354):65-70. PMC: 3131495. DOI: 10.1038/nature10236. View

15.

Schneider E, Schnell D, Papa D, Seifert R . High constitutive activity and a G-protein-independent high-affinity state of the human histamine H(4)-receptor. Biochemistry. 2009; 48(6):1424-38. DOI: 10.1021/bi802050d. View

16.

Rossbach K, Wendorff S, Sander K, Stark H, Gutzmer R, Werfel T . Histamine H4 receptor antagonism reduces hapten-induced scratching behaviour but not inflammation. Exp Dermatol. 2008; 18(1):57-63. DOI: 10.1111/j.1600-0625.2008.00762.x. View

17.

Shin N, Coates E, Murgolo N, Morse K, Bayne M, Strader C . Molecular modeling and site-specific mutagenesis of the histamine-binding site of the histamine H4 receptor. Mol Pharmacol. 2002; 62(1):38-47. DOI: 10.1124/mol.62.1.38. View

18.

de Esch I, Thurmond R, Jongejan A, Leurs R . The histamine H4 receptor as a new therapeutic target for inflammation. Trends Pharmacol Sci. 2005; 26(9):462-9. DOI: 10.1016/j.tips.2005.07.002. View

19.

Zhu Y, Michalovich D, Wu H, Tan K, Dytko G, Mannan I . Cloning, expression, and pharmacological characterization of a novel human histamine receptor. Mol Pharmacol. 2001; 59(3):434-41. DOI: 10.1124/mol.59.3.434. View

20.

Ling P, Ngo K, Nguyen S, Thurmond R, Edwards J, Karlsson L . Histamine H4 receptor mediates eosinophil chemotaxis with cell shape change and adhesion molecule upregulation. Br J Pharmacol. 2004; 142(1):161-71. PMC: 1574921. DOI: 10.1038/sj.bjp.0705729. View