title: Phenylalanine 169 in the second extracellular loop of the human histamine H4 receptor is responsible for the difference in agonist binding between human and mouse H4 receptors.
abstract: Using the natural variation in histamine H(4) receptor protein sequence, we tried to identify amino acids involved in the binding of H(4) receptor agonists. To this end, we constructed a variety of chimeric human-mouse H(4) receptor proteins to localize the domain responsible for the observed pharmacological differences between human and mouse H(4) receptors in the binding of H(4) receptor agonists, such as histamine, clozapine, and VUF 8430 [S-(2-guanidylethyl)-isothiourea]. After identification of a domain between the top of transmembrane domain 4 and the top of transmembrane domain 5 as being responsible for the differences in agonist affinity between human and mouse H(4)Rs, detailed site-directed mutagenesis studies were performed. These studies identified Phe(169) in the second extracellular loop as the single amino acid responsible for the differences in agonist affinity between the human and mouse H(4)Rs. Phe(169) is part of a Phe-Phe motif, which is also present in the recently crystallized beta(2)-adrenergic receptor. These results point to an important role of the second extracellular loop in the agonist binding to the H(4) receptor and provide a molecular explanation for the species difference between human and mouse H(4) receptors.
authors.affiliation: Leiden/Amsterdam Center for Drug Research, Division of Medicinal Chemistry, Faculty of Science, Vrije Universiteit Amsterdam, De Boelelaan 1083, 1081 HV Amsterdam, The Netherlands. r.leurs@few.vu.nl.
authors.forename: Herman D
authors.initials: HD
authors.lastname: Lim
authors.forename: Aldo
authors.initials: A
authors.lastname: Jongejan
authors.forename: Remko A
authors.initials: RA
authors.lastname: Bakker
authors.forename: Eric
authors.initials: E
authors.lastname: Haaksma
authors.forename: Iwan J P
authors.initials: IJ
authors.lastname: de Esch
authors.forename: Rob
authors.initials: R
authors.lastname: Leurs
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002460:Cell Line; D006632:Histamine / Q000378:metabolism; D006801:Humans; D007211:Indoles / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D010649:Phenylalanine; D010879:Piperazines / Q000378:metabolism; D018343:Receptors, Adrenergic, beta-2 / Q000737:chemistry; D043562:Receptors, G-Protein-Coupled / Q000819:agonists / Q000737:chemistry*; D011968:Receptors, Histamine / Q000737:chemistry*; D000074040:Receptors, Histamine H4; D013045:Species Specificity
chemical_list: C418607:HRH4 protein, human; C498829:Hrh4 protein, mouse; D007211:Indoles; D010879:Piperazines; D018343:Receptors, Adrenergic, beta-2; D043562:Receptors, G-Protein-Coupled; D011968:Receptors, Histamine; D000074040:Receptors, Histamine H4; D010649:Phenylalanine; C484309:1-((5-chloro-1H-indol-2-yl)carbonyl)-4-methylpiperazine; D006632:Histamine