A high affinity, highly selective ligand for the delta opioid receptor: [3H]-[D-PEN2, pCl-PHE4, D-PEN5]enkephalin

Linda K. Vaughn, Richard J. Knapp, Geza Toth, Y. P. Wan, Victor J Hruby, Henry I. Yamamura

Research output: Contribution to journalArticle

86 Citations (Scopus)

Abstract

Binding characteristics of a new, conformationally constrained, halogenated enkephalin analogue, [3H]-[D-penicillamine2, pCl-Phe4, D-penicillamine5]enkephalin ([3H]pCl-DPDPE), were determined using homogenized rat brain tissue. Saturation binding studies at 25°C determined a dissociation constant (Kd) of 328 ± 27 pM and a receptor density (Bmax) of 87.2 ± 4.2 fmol/mg protein. Kinetic studies demonstrated biphasic association for [3H]pCl-DPDPE, with association rate constants of 5.05 × 108 ± 2.5 × 108 and 0.147 ± 108 ± 0.014 × 108 M-1 min-1. Dissociation was monophasic with a dissociation rate constant of 2.96 × 10-3 ± 0.25 × 10-1 min-1. The average Kd values determined by these kinetic studies were 8.4 ± 2.7 pM and 201 ± 4 pM. Competitive inhibition studies demonstrated that [3H]pCl-DPDPE has excellent selectively for the delta opioid receptor. [3H]pCl-DPDPE binding was inhibited by low concentrations of ligands selective for delta opioid receptor relative to the concentrations required by ligands selective for mu and kappa sites. These data show that [3H]pCl-DPDPE is a highly selective, high affinity ligand which should be useful in characterizing the delta opioid receptor.

Original languageEnglish (US)
Pages (from-to)1001-1008
Number of pages8
JournalLife Sciences
Volume45
Issue number11
DOIs
StatePublished - 1989

Fingerprint

delta Opioid Receptor
Ligands
Rate constants
Association reactions
Kinetics
Enkephalins
Rats
Brain
Ala(2)-enkephalinamide-met
Pen(2,5)-4-chloro-Phe(4)-enkephalin
Tissue
Proteins

ASJC Scopus subject areas

  • Pharmacology

Cite this

A high affinity, highly selective ligand for the delta opioid receptor : [3H]-[D-PEN2, pCl-PHE4, D-PEN5]enkephalin. / Vaughn, Linda K.; Knapp, Richard J.; Toth, Geza; Wan, Y. P.; Hruby, Victor J; Yamamura, Henry I.

In: Life Sciences, Vol. 45, No. 11, 1989, p. 1001-1008.

Research output: Contribution to journalArticle

Vaughn, Linda K. ; Knapp, Richard J. ; Toth, Geza ; Wan, Y. P. ; Hruby, Victor J ; Yamamura, Henry I. / A high affinity, highly selective ligand for the delta opioid receptor : [3H]-[D-PEN2, pCl-PHE4, D-PEN5]enkephalin. In: Life Sciences. 1989 ; Vol. 45, No. 11. pp. 1001-1008.
@article{94378a7140ee4b06909fdf797ba0e70e,
title = "A high affinity, highly selective ligand for the delta opioid receptor: [3H]-[D-PEN2, pCl-PHE4, D-PEN5]enkephalin",
abstract = "Binding characteristics of a new, conformationally constrained, halogenated enkephalin analogue, [3H]-[D-penicillamine2, pCl-Phe4, D-penicillamine5]enkephalin ([3H]pCl-DPDPE), were determined using homogenized rat brain tissue. Saturation binding studies at 25°C determined a dissociation constant (Kd) of 328 ± 27 pM and a receptor density (Bmax) of 87.2 ± 4.2 fmol/mg protein. Kinetic studies demonstrated biphasic association for [3H]pCl-DPDPE, with association rate constants of 5.05 × 108 ± 2.5 × 108 and 0.147 ± 108 ± 0.014 × 108 M-1 min-1. Dissociation was monophasic with a dissociation rate constant of 2.96 × 10-3 ± 0.25 × 10-1 min-1. The average Kd values determined by these kinetic studies were 8.4 ± 2.7 pM and 201 ± 4 pM. Competitive inhibition studies demonstrated that [3H]pCl-DPDPE has excellent selectively for the delta opioid receptor. [3H]pCl-DPDPE binding was inhibited by low concentrations of ligands selective for delta opioid receptor relative to the concentrations required by ligands selective for mu and kappa sites. These data show that [3H]pCl-DPDPE is a highly selective, high affinity ligand which should be useful in characterizing the delta opioid receptor.",
author = "Vaughn, {Linda K.} and Knapp, {Richard J.} and Geza Toth and Wan, {Y. P.} and Hruby, {Victor J} and Yamamura, {Henry I.}",
year = "1989",
doi = "10.1016/0024-3205(89)90154-9",
language = "English (US)",
volume = "45",
pages = "1001--1008",
journal = "Life Sciences",
issn = "0024-3205",
publisher = "Elsevier Inc.",
number = "11",

}

TY - JOUR

T1 - A high affinity, highly selective ligand for the delta opioid receptor

T2 - [3H]-[D-PEN2, pCl-PHE4, D-PEN5]enkephalin

AU - Vaughn, Linda K.

AU - Knapp, Richard J.

AU - Toth, Geza

AU - Wan, Y. P.

AU - Hruby, Victor J

AU - Yamamura, Henry I.

PY - 1989

Y1 - 1989

N2 - Binding characteristics of a new, conformationally constrained, halogenated enkephalin analogue, [3H]-[D-penicillamine2, pCl-Phe4, D-penicillamine5]enkephalin ([3H]pCl-DPDPE), were determined using homogenized rat brain tissue. Saturation binding studies at 25°C determined a dissociation constant (Kd) of 328 ± 27 pM and a receptor density (Bmax) of 87.2 ± 4.2 fmol/mg protein. Kinetic studies demonstrated biphasic association for [3H]pCl-DPDPE, with association rate constants of 5.05 × 108 ± 2.5 × 108 and 0.147 ± 108 ± 0.014 × 108 M-1 min-1. Dissociation was monophasic with a dissociation rate constant of 2.96 × 10-3 ± 0.25 × 10-1 min-1. The average Kd values determined by these kinetic studies were 8.4 ± 2.7 pM and 201 ± 4 pM. Competitive inhibition studies demonstrated that [3H]pCl-DPDPE has excellent selectively for the delta opioid receptor. [3H]pCl-DPDPE binding was inhibited by low concentrations of ligands selective for delta opioid receptor relative to the concentrations required by ligands selective for mu and kappa sites. These data show that [3H]pCl-DPDPE is a highly selective, high affinity ligand which should be useful in characterizing the delta opioid receptor.

AB - Binding characteristics of a new, conformationally constrained, halogenated enkephalin analogue, [3H]-[D-penicillamine2, pCl-Phe4, D-penicillamine5]enkephalin ([3H]pCl-DPDPE), were determined using homogenized rat brain tissue. Saturation binding studies at 25°C determined a dissociation constant (Kd) of 328 ± 27 pM and a receptor density (Bmax) of 87.2 ± 4.2 fmol/mg protein. Kinetic studies demonstrated biphasic association for [3H]pCl-DPDPE, with association rate constants of 5.05 × 108 ± 2.5 × 108 and 0.147 ± 108 ± 0.014 × 108 M-1 min-1. Dissociation was monophasic with a dissociation rate constant of 2.96 × 10-3 ± 0.25 × 10-1 min-1. The average Kd values determined by these kinetic studies were 8.4 ± 2.7 pM and 201 ± 4 pM. Competitive inhibition studies demonstrated that [3H]pCl-DPDPE has excellent selectively for the delta opioid receptor. [3H]pCl-DPDPE binding was inhibited by low concentrations of ligands selective for delta opioid receptor relative to the concentrations required by ligands selective for mu and kappa sites. These data show that [3H]pCl-DPDPE is a highly selective, high affinity ligand which should be useful in characterizing the delta opioid receptor.

UR - http://www.scopus.com/inward/record.url?scp=0024423167&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0024423167&partnerID=8YFLogxK

U2 - 10.1016/0024-3205(89)90154-9

DO - 10.1016/0024-3205(89)90154-9

M3 - Article

C2 - 2552241

AN - SCOPUS:0024423167

VL - 45

SP - 1001

EP - 1008

JO - Life Sciences

JF - Life Sciences

SN - 0024-3205

IS - 11

ER -