Synergistic effect of basic residues at positions 14-15 of nociceptin on binding affinity and receptor activation

Kazushi Okada, Kaname Isozaki, Jinglan Li, Ayami Matsushima, Takeru Nose, Tommaso Costa, Yasuyuki Shimohigashi

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

Nociceptin is an endogenous ligand that activates a G protein-coupled receptor ORL1 and contains two indispensable Arg-Lys (RK) dipeptide units at positions 8-9 and 12-13. By replacing an additional RK unit at positions 6-7, 10-11, 14-15, or 16-17, of the peptide we have identified the analog, [RK14-15]nociceptin as a superagonist. In fact, this peptide exhibits 3-fold higher binding affinity and 17-fold greater potency in a functional GTPγS-binding assay compared to wild-type nociceptin. Here, we have further investigated the role of basic residues in position 14-15. The replacement of three other possible basic dipeptides, KR, RR, and KK, into nociceptin at positions 14-15 resulted in similar enhancements of binding affinity (3-5-fold) and biological potency (10-12-fold in the GTPγS assay). However, when only a single basic residue (Arg or Lys) was replaced in either position 14 or 15, all the resulting analogs showed moderate enhancements of binding and biological activity (2-4-fold in both). These results indicate that the addition of basic charges in positions 14 and 15 enhance in a synergistic fashion the interaction of nociceptin with the receptor and only the simultaneous presence of two adjacent basic residues yields an optimal effect. This suggests that specific electrostatic interactions between both amino acids present in 14-15 and corresponding residues in the receptor are responsible for the enhancement of nociceptin activity.

Original languageEnglish
Pages (from-to)9261-9267
Number of pages7
JournalBioorganic and Medicinal Chemistry
Volume16
Issue number20
DOIs
Publication statusPublished - Oct 15 2008
Externally publishedYes

Fingerprint

Chemical activation
Dipeptides
arginyllysine
Assays
Peptides
G-Protein-Coupled Receptors
Static Electricity
Coulomb interactions
Bioactivity
nociceptin
Ligands
Amino Acids

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmaceutical Science
  • Drug Discovery
  • Clinical Biochemistry
  • Organic Chemistry

Cite this

Synergistic effect of basic residues at positions 14-15 of nociceptin on binding affinity and receptor activation. / Okada, Kazushi; Isozaki, Kaname; Li, Jinglan; Matsushima, Ayami; Nose, Takeru; Costa, Tommaso; Shimohigashi, Yasuyuki.

In: Bioorganic and Medicinal Chemistry, Vol. 16, No. 20, 15.10.2008, p. 9261-9267.

Research output: Contribution to journalArticle

Okada, Kazushi ; Isozaki, Kaname ; Li, Jinglan ; Matsushima, Ayami ; Nose, Takeru ; Costa, Tommaso ; Shimohigashi, Yasuyuki. / Synergistic effect of basic residues at positions 14-15 of nociceptin on binding affinity and receptor activation. In: Bioorganic and Medicinal Chemistry. 2008 ; Vol. 16, No. 20. pp. 9261-9267.
@article{13db83b0c3e44582b5ffa553d2643c7c,
title = "Synergistic effect of basic residues at positions 14-15 of nociceptin on binding affinity and receptor activation",
abstract = "Nociceptin is an endogenous ligand that activates a G protein-coupled receptor ORL1 and contains two indispensable Arg-Lys (RK) dipeptide units at positions 8-9 and 12-13. By replacing an additional RK unit at positions 6-7, 10-11, 14-15, or 16-17, of the peptide we have identified the analog, [RK14-15]nociceptin as a superagonist. In fact, this peptide exhibits 3-fold higher binding affinity and 17-fold greater potency in a functional GTPγS-binding assay compared to wild-type nociceptin. Here, we have further investigated the role of basic residues in position 14-15. The replacement of three other possible basic dipeptides, KR, RR, and KK, into nociceptin at positions 14-15 resulted in similar enhancements of binding affinity (3-5-fold) and biological potency (10-12-fold in the GTPγS assay). However, when only a single basic residue (Arg or Lys) was replaced in either position 14 or 15, all the resulting analogs showed moderate enhancements of binding and biological activity (2-4-fold in both). These results indicate that the addition of basic charges in positions 14 and 15 enhance in a synergistic fashion the interaction of nociceptin with the receptor and only the simultaneous presence of two adjacent basic residues yields an optimal effect. This suggests that specific electrostatic interactions between both amino acids present in 14-15 and corresponding residues in the receptor are responsible for the enhancement of nociceptin activity.",
author = "Kazushi Okada and Kaname Isozaki and Jinglan Li and Ayami Matsushima and Takeru Nose and Tommaso Costa and Yasuyuki Shimohigashi",
year = "2008",
month = "10",
day = "15",
doi = "10.1016/j.bmc.2008.09.014",
language = "English",
volume = "16",
pages = "9261--9267",
journal = "Bioorganic and Medicinal Chemistry",
issn = "0968-0896",
publisher = "Elsevier Limited",
number = "20",

}

TY - JOUR

T1 - Synergistic effect of basic residues at positions 14-15 of nociceptin on binding affinity and receptor activation

AU - Okada, Kazushi

AU - Isozaki, Kaname

AU - Li, Jinglan

AU - Matsushima, Ayami

AU - Nose, Takeru

AU - Costa, Tommaso

AU - Shimohigashi, Yasuyuki

PY - 2008/10/15

Y1 - 2008/10/15

N2 - Nociceptin is an endogenous ligand that activates a G protein-coupled receptor ORL1 and contains two indispensable Arg-Lys (RK) dipeptide units at positions 8-9 and 12-13. By replacing an additional RK unit at positions 6-7, 10-11, 14-15, or 16-17, of the peptide we have identified the analog, [RK14-15]nociceptin as a superagonist. In fact, this peptide exhibits 3-fold higher binding affinity and 17-fold greater potency in a functional GTPγS-binding assay compared to wild-type nociceptin. Here, we have further investigated the role of basic residues in position 14-15. The replacement of three other possible basic dipeptides, KR, RR, and KK, into nociceptin at positions 14-15 resulted in similar enhancements of binding affinity (3-5-fold) and biological potency (10-12-fold in the GTPγS assay). However, when only a single basic residue (Arg or Lys) was replaced in either position 14 or 15, all the resulting analogs showed moderate enhancements of binding and biological activity (2-4-fold in both). These results indicate that the addition of basic charges in positions 14 and 15 enhance in a synergistic fashion the interaction of nociceptin with the receptor and only the simultaneous presence of two adjacent basic residues yields an optimal effect. This suggests that specific electrostatic interactions between both amino acids present in 14-15 and corresponding residues in the receptor are responsible for the enhancement of nociceptin activity.

AB - Nociceptin is an endogenous ligand that activates a G protein-coupled receptor ORL1 and contains two indispensable Arg-Lys (RK) dipeptide units at positions 8-9 and 12-13. By replacing an additional RK unit at positions 6-7, 10-11, 14-15, or 16-17, of the peptide we have identified the analog, [RK14-15]nociceptin as a superagonist. In fact, this peptide exhibits 3-fold higher binding affinity and 17-fold greater potency in a functional GTPγS-binding assay compared to wild-type nociceptin. Here, we have further investigated the role of basic residues in position 14-15. The replacement of three other possible basic dipeptides, KR, RR, and KK, into nociceptin at positions 14-15 resulted in similar enhancements of binding affinity (3-5-fold) and biological potency (10-12-fold in the GTPγS assay). However, when only a single basic residue (Arg or Lys) was replaced in either position 14 or 15, all the resulting analogs showed moderate enhancements of binding and biological activity (2-4-fold in both). These results indicate that the addition of basic charges in positions 14 and 15 enhance in a synergistic fashion the interaction of nociceptin with the receptor and only the simultaneous presence of two adjacent basic residues yields an optimal effect. This suggests that specific electrostatic interactions between both amino acids present in 14-15 and corresponding residues in the receptor are responsible for the enhancement of nociceptin activity.

UR - http://www.scopus.com/inward/record.url?scp=53449086268&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=53449086268&partnerID=8YFLogxK

U2 - 10.1016/j.bmc.2008.09.014

DO - 10.1016/j.bmc.2008.09.014

M3 - Article

C2 - 18818087

AN - SCOPUS:53449086268

VL - 16

SP - 9261

EP - 9267

JO - Bioorganic and Medicinal Chemistry

JF - Bioorganic and Medicinal Chemistry

SN - 0968-0896

IS - 20

ER -