Molecular basis for agonism in the BB3 receptor: an epitope located on the interface of transmembrane-III, -VI, and -VII

F Gbahou; B Holst; T W Schwartz

doi:10.1124/jpet.109.162131

Molecular basis for agonism in the BB3 receptor: an epitope located on the interface of transmembrane-III, -VI, and -VII

F Gbahou, B Holst, T W Schwartz

Institut for Neurovidenskab

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › peer review

17 Citationer (Scopus)

Abstract

Udgivelsesdato: 2010-Apr

Originalsprog	Engelsk
Tidsskrift	Journal of Pharmacology and Experimental Therapeutics
Vol/bind	333
Udgave nummer	1
Sider (fra-til)	51-9
Antal sider	8
ISSN	0022-3565
DOI	https://doi.org/10.1124/jpet.109.162131
Status	Udgivet - 2010

Bibliografisk note

Keywords: Binding Sites; Bombesin; Cell Line; Epitopes; Humans; Ligands; Models, Molecular; Mutation; Oligopeptides; Peptide Fragments; Receptors, Bombesin; Signal Transduction; Structure-Activity Relationship

Adgang til dokumentet

10.1124/jpet.109.162131

Citationsformater

@article{d2402a40b42e11df825b000ea68e967b,

title = "Molecular basis for agonism in the BB3 receptor: an epitope located on the interface of transmembrane-III, -VI, and -VII",

abstract = "Epitopes determining the agonist property of two structurally distinct selective ligands for the human bombesin receptor subtype 3 (BB3), [D-Tyr6,(R)-Apa11,Phe13, Nle14]-bombesin(6-14) (Pep-1) and Ac-Phe-Trp-Ala-His(TauBzl)-Nip-Gly-Arg-NH2 (Pep-2), were mapped through systematic mutagenesis of the main ligand-binding pocket of the receptor. The mutational map for the smaller Pep-2 spanned the entire binding pocket of the BB3 receptor. In contrast, the much fewer mutational hits for the larger Pep-1 were confined to the center of the pocket, i.e., the opposing faces of the extracellular segments of transmembrane (TM)-III, TM-VI, and TM-VII. All the residues, which upon mutation affected Pep-1, were also hits for Pep-2 and included those that were most essential for the function of Pep-2: LeuIII:04 (Leu(123)), TyrVI:16 (Tyr(291)), and ArgVII:06 (Arg(316)). The BB3 receptor was found to signal with 12% ligand-independent activity that was strongly influenced both positively and negatively by several mutations in the binding pocket. The substitutions, which decreased the constitutive signaling, included not only the major mutational hits for the peptide agonists but also mutations more superficially located in the receptor. It is concluded that activation of the BB3 receptor is dependent upon an epitope in the main ligand-binding pocket at the interface between TM-III, TM-VI, and TM-VII that corresponds to the site where, for example, activating metal ion sites have been constructed previously in 7TM receptors.",

author = "F Gbahou and B Holst and Schwartz, {T W}",

note = "Keywords: Binding Sites; Bombesin; Cell Line; Epitopes; Humans; Ligands; Models, Molecular; Mutation; Oligopeptides; Peptide Fragments; Receptors, Bombesin; Signal Transduction; Structure-Activity Relationship",

year = "2010",

doi = "10.1124/jpet.109.162131",

language = "English",

volume = "333",

pages = "51--9",

journal = "Journal of Pharmacology and Experimental Therapeutics",

issn = "0022-3565",

publisher = "American Society for Pharmacology and Experimental Therapeutics",

number = "1",

}

TY - JOUR

T1 - Molecular basis for agonism in the BB3 receptor: an epitope located on the interface of transmembrane-III, -VI, and -VII

AU - Gbahou, F

AU - Holst, B

AU - Schwartz, T W

N1 - Keywords: Binding Sites; Bombesin; Cell Line; Epitopes; Humans; Ligands; Models, Molecular; Mutation; Oligopeptides; Peptide Fragments; Receptors, Bombesin; Signal Transduction; Structure-Activity Relationship

PY - 2010

Y1 - 2010

N2 - Epitopes determining the agonist property of two structurally distinct selective ligands for the human bombesin receptor subtype 3 (BB3), [D-Tyr6,(R)-Apa11,Phe13, Nle14]-bombesin(6-14) (Pep-1) and Ac-Phe-Trp-Ala-His(TauBzl)-Nip-Gly-Arg-NH2 (Pep-2), were mapped through systematic mutagenesis of the main ligand-binding pocket of the receptor. The mutational map for the smaller Pep-2 spanned the entire binding pocket of the BB3 receptor. In contrast, the much fewer mutational hits for the larger Pep-1 were confined to the center of the pocket, i.e., the opposing faces of the extracellular segments of transmembrane (TM)-III, TM-VI, and TM-VII. All the residues, which upon mutation affected Pep-1, were also hits for Pep-2 and included those that were most essential for the function of Pep-2: LeuIII:04 (Leu(123)), TyrVI:16 (Tyr(291)), and ArgVII:06 (Arg(316)). The BB3 receptor was found to signal with 12% ligand-independent activity that was strongly influenced both positively and negatively by several mutations in the binding pocket. The substitutions, which decreased the constitutive signaling, included not only the major mutational hits for the peptide agonists but also mutations more superficially located in the receptor. It is concluded that activation of the BB3 receptor is dependent upon an epitope in the main ligand-binding pocket at the interface between TM-III, TM-VI, and TM-VII that corresponds to the site where, for example, activating metal ion sites have been constructed previously in 7TM receptors.

AB - Epitopes determining the agonist property of two structurally distinct selective ligands for the human bombesin receptor subtype 3 (BB3), [D-Tyr6,(R)-Apa11,Phe13, Nle14]-bombesin(6-14) (Pep-1) and Ac-Phe-Trp-Ala-His(TauBzl)-Nip-Gly-Arg-NH2 (Pep-2), were mapped through systematic mutagenesis of the main ligand-binding pocket of the receptor. The mutational map for the smaller Pep-2 spanned the entire binding pocket of the BB3 receptor. In contrast, the much fewer mutational hits for the larger Pep-1 were confined to the center of the pocket, i.e., the opposing faces of the extracellular segments of transmembrane (TM)-III, TM-VI, and TM-VII. All the residues, which upon mutation affected Pep-1, were also hits for Pep-2 and included those that were most essential for the function of Pep-2: LeuIII:04 (Leu(123)), TyrVI:16 (Tyr(291)), and ArgVII:06 (Arg(316)). The BB3 receptor was found to signal with 12% ligand-independent activity that was strongly influenced both positively and negatively by several mutations in the binding pocket. The substitutions, which decreased the constitutive signaling, included not only the major mutational hits for the peptide agonists but also mutations more superficially located in the receptor. It is concluded that activation of the BB3 receptor is dependent upon an epitope in the main ligand-binding pocket at the interface between TM-III, TM-VI, and TM-VII that corresponds to the site where, for example, activating metal ion sites have been constructed previously in 7TM receptors.

U2 - 10.1124/jpet.109.162131

DO - 10.1124/jpet.109.162131

M3 - Journal article

C2 - 20065020

SN - 0022-3565

VL - 333

SP - 51

EP - 59

JO - Journal of Pharmacology and Experimental Therapeutics

JF - Journal of Pharmacology and Experimental Therapeutics

IS - 1

ER -