Unique interaction pattern for a functionally biased ghrelin receptor agonist

Bjørn Behrens Sivertsen; Manja Lang; Thomas M. Frimurer; Nicholas D. Holliday; Anders Bach; Sylvia Els; Maja Storm Engelstoft; Pia Steen Petersen; Andreas Nygaard Madsen; Thue W. Schwartz; Annette G. Beck-Sickinger; Birgitte Holst

doi:10.1074/jbc.M110.173237

Unique interaction pattern for a functionally biased ghrelin receptor agonist

Bjørn Behrens Sivertsen, Manja Lang, Thomas M. Frimurer, Nicholas D. Holliday, Anders Bach, Sylvia Els, Maja Storm Engelstoft, Pia Steen Petersen, Andreas Nygaard Madsen, Thue W. Schwartz, Annette G. Beck-Sickinger, Birgitte Holst

38 Citations (Scopus)

Abstract

Based on the conformationally constrained D-Trp-Phe-D-Trp (wFw) core of the prototype inverse agonist [D-Arg¹,D-Phe⁵,D-Trp ^7,9, Leu¹¹]substance P, a series of novel, small, peptide-mimetic agonists for the ghrelin receptor were generated. By using various simple, ring-constrained spacers connecting the D-Trp-Phe-D-Trp motif with the important C-terminal carboxyamide group, 40 nM agonism potency was obtained and also in one case (wFw-Isn-NH₂, where Isn is isonipecotic acid) ∼80% efficacy. However, in contrast to all previously reported ghrelin receptor agonists, the piperidine-constrained wFw-Isn-NH₂ was found to be a functionally biased agonist. Thus, wFw-Isn-NH₂ mediated potent and efficacious signaling through the Gα_q and ERK1/2 signaling pathways, but in contrast to all previous ghrelin receptor agonists it did not signal through the serum response element, conceivably the Gα_12/13 pathway. The recognition pattern of wFw-Isn-NH ₂ with the ghrelin receptor also differed significantly from that of all previously characterized unbiased agonists. Most importantly, wFw-Isn-NH₂ was not dependent on GluIII:09 (Glu3.33), which otherwise is an obligatory TM III anchor point residue for ghrelin agonists. Molecular modeling and docking experiments indicated that wFw-Isn-NH₂ binds in the classical agonist binding site between the extracellular segments of TMs III, VI, and VII, interacting closely with the aromatic cluster between TMs VI and VII, but that it does so in an opposite orientation as compared with, for example, the wFw peptide agonists. It is concluded that the novel peptide-mimetic ligand wFw-Isn-NH₂ is a biased ghrelin receptor agonist and that the selective signaling pattern presumably is due to its unique receptor recognition pattern lacking interaction with key residues especially in TM III.

Original language	English
Journal	The Journal of Biological Chemistry
Volume	286
Issue number	23
Pages (from-to)	20845-20860
Number of pages	16
ISSN	1083-351X
DOIs	https://doi.org/10.1074/jbc.M110.173237
Publication status	Published - 10 Jun 2011

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@article{64992d7adb904db2809c740335efc150,

title = "Unique interaction pattern for a functionally biased ghrelin receptor agonist",

abstract = "Based on the conformationally constrained D-Trp-Phe-D-Trp (wFw) core of the prototype inverse agonist [D-Arg1,D-Phe5,D-Trp 7,9, Leu11]substance P, a series of novel, small, peptide-mimetic agonists for the ghrelin receptor were generated. By using various simple, ring-constrained spacers connecting the D-Trp-Phe-D-Trp motif with the important C-terminal carboxyamide group, 40 nM agonism potency was obtained and also in one case (wFw-Isn-NH2, where Isn is isonipecotic acid) ∼80% efficacy. However, in contrast to all previously reported ghrelin receptor agonists, the piperidine-constrained wFw-Isn-NH2 was found to be a functionally biased agonist. Thus, wFw-Isn-NH2 mediated potent and efficacious signaling through the Gαq and ERK1/2 signaling pathways, but in contrast to all previous ghrelin receptor agonists it did not signal through the serum response element, conceivably the Gα12/13 pathway. The recognition pattern of wFw-Isn-NH 2 with the ghrelin receptor also differed significantly from that of all previously characterized unbiased agonists. Most importantly, wFw-Isn-NH2 was not dependent on GluIII:09 (Glu3.33), which otherwise is an obligatory TM III anchor point residue for ghrelin agonists. Molecular modeling and docking experiments indicated that wFw-Isn-NH2 binds in the classical agonist binding site between the extracellular segments of TMs III, VI, and VII, interacting closely with the aromatic cluster between TMs VI and VII, but that it does so in an opposite orientation as compared with, for example, the wFw peptide agonists. It is concluded that the novel peptide-mimetic ligand wFw-Isn-NH2 is a biased ghrelin receptor agonist and that the selective signaling pattern presumably is due to its unique receptor recognition pattern lacking interaction with key residues especially in TM III.",

author = "Sivertsen, {Bj{\o}rn Behrens} and Manja Lang and Frimurer, {Thomas M.} and Holliday, {Nicholas D.} and Anders Bach and Sylvia Els and Engelstoft, {Maja Storm} and Petersen, {Pia Steen} and Madsen, {Andreas Nygaard} and Schwartz, {Thue W.} and Beck-Sickinger, {Annette G.} and Birgitte Holst",

year = "2011",

month = jun,

day = "10",

doi = "10.1074/jbc.M110.173237",

language = "English",

volume = "286",

pages = "20845--20860",

journal = "Journal of Biological Chemistry",

issn = "0021-9258",

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T1 - Unique interaction pattern for a functionally biased ghrelin receptor agonist

AU - Sivertsen, Bjørn Behrens

AU - Lang, Manja

AU - Frimurer, Thomas M.

AU - Holliday, Nicholas D.

AU - Bach, Anders

AU - Els, Sylvia

AU - Engelstoft, Maja Storm

AU - Petersen, Pia Steen

AU - Madsen, Andreas Nygaard

AU - Schwartz, Thue W.

AU - Beck-Sickinger, Annette G.

AU - Holst, Birgitte

PY - 2011/6/10

Y1 - 2011/6/10

N2 - Based on the conformationally constrained D-Trp-Phe-D-Trp (wFw) core of the prototype inverse agonist [D-Arg1,D-Phe5,D-Trp 7,9, Leu11]substance P, a series of novel, small, peptide-mimetic agonists for the ghrelin receptor were generated. By using various simple, ring-constrained spacers connecting the D-Trp-Phe-D-Trp motif with the important C-terminal carboxyamide group, 40 nM agonism potency was obtained and also in one case (wFw-Isn-NH2, where Isn is isonipecotic acid) ∼80% efficacy. However, in contrast to all previously reported ghrelin receptor agonists, the piperidine-constrained wFw-Isn-NH2 was found to be a functionally biased agonist. Thus, wFw-Isn-NH2 mediated potent and efficacious signaling through the Gαq and ERK1/2 signaling pathways, but in contrast to all previous ghrelin receptor agonists it did not signal through the serum response element, conceivably the Gα12/13 pathway. The recognition pattern of wFw-Isn-NH 2 with the ghrelin receptor also differed significantly from that of all previously characterized unbiased agonists. Most importantly, wFw-Isn-NH2 was not dependent on GluIII:09 (Glu3.33), which otherwise is an obligatory TM III anchor point residue for ghrelin agonists. Molecular modeling and docking experiments indicated that wFw-Isn-NH2 binds in the classical agonist binding site between the extracellular segments of TMs III, VI, and VII, interacting closely with the aromatic cluster between TMs VI and VII, but that it does so in an opposite orientation as compared with, for example, the wFw peptide agonists. It is concluded that the novel peptide-mimetic ligand wFw-Isn-NH2 is a biased ghrelin receptor agonist and that the selective signaling pattern presumably is due to its unique receptor recognition pattern lacking interaction with key residues especially in TM III.

AB - Based on the conformationally constrained D-Trp-Phe-D-Trp (wFw) core of the prototype inverse agonist [D-Arg1,D-Phe5,D-Trp 7,9, Leu11]substance P, a series of novel, small, peptide-mimetic agonists for the ghrelin receptor were generated. By using various simple, ring-constrained spacers connecting the D-Trp-Phe-D-Trp motif with the important C-terminal carboxyamide group, 40 nM agonism potency was obtained and also in one case (wFw-Isn-NH2, where Isn is isonipecotic acid) ∼80% efficacy. However, in contrast to all previously reported ghrelin receptor agonists, the piperidine-constrained wFw-Isn-NH2 was found to be a functionally biased agonist. Thus, wFw-Isn-NH2 mediated potent and efficacious signaling through the Gαq and ERK1/2 signaling pathways, but in contrast to all previous ghrelin receptor agonists it did not signal through the serum response element, conceivably the Gα12/13 pathway. The recognition pattern of wFw-Isn-NH 2 with the ghrelin receptor also differed significantly from that of all previously characterized unbiased agonists. Most importantly, wFw-Isn-NH2 was not dependent on GluIII:09 (Glu3.33), which otherwise is an obligatory TM III anchor point residue for ghrelin agonists. Molecular modeling and docking experiments indicated that wFw-Isn-NH2 binds in the classical agonist binding site between the extracellular segments of TMs III, VI, and VII, interacting closely with the aromatic cluster between TMs VI and VII, but that it does so in an opposite orientation as compared with, for example, the wFw peptide agonists. It is concluded that the novel peptide-mimetic ligand wFw-Isn-NH2 is a biased ghrelin receptor agonist and that the selective signaling pattern presumably is due to its unique receptor recognition pattern lacking interaction with key residues especially in TM III.

U2 - 10.1074/jbc.M110.173237

DO - 10.1074/jbc.M110.173237

M3 - Journal article

C2 - 21402696

SN - 0021-9258

VL - 286

SP - 20845

EP - 20860

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

IS - 23

ER -

Unique interaction pattern for a functionally biased ghrelin receptor agonist

Abstract

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