Partial characterization of the specific binding of 3H-N-allylnormetazocine (SKF 10047) to rat brain membranes

K. M. Johnson

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The specific binding of 3H(±)-N-allylnormetazocine (3H(±)-SKF 10047) to rat brain membranes was defined as that which could be displaced by 100μM (+)SKF 10047. This binding was found to be saturable and Scatchard analysis indicated the presence of three binding components with differing apparent K(D) and B(max) values. These sites varied considerably in their sensitivity to the effects of heat, trypsin, and ionic strength, with the low affinity site (K(D) = 245 nM) being least sensitive to elevated temperature and trypsinization and most sensitive to elevation of the ionic strength of the incubation buffer. A modest degree of selectivity could be attained for each of the three sites by varying the concentration of 3H(±)-SKF 10047, e.g. at 160 nM 3H(±)-SKF 10047 approximately 75% of the total binding was calculated to be due to the low-affinity site. Using this technique it was found that both isomers of SKF 10047 were equi-effective in displacing binding from the low-affinity site, but (-)SKF 10047 was 150 times more potent than (+)SKF 10047 at the high-affinity site. Also, naloxone was very potent in inhibiting binding to the high-affinity site (IC50 = 4.7 nM), but very weak at the low-affinity site (IC50 = 100 μM). Phencyclidine (PCP) and several of its derivatives were found to have potencies similar to those of SKF 10047 in inhibiting binding to the low-affinity site. These data suggest that the low-affinity binding site for this prototypic sigma opiate may be analogous to the putative PCP receptor.

Original languageEnglish (US)
Pages (from-to)33-47
Number of pages15
JournalResearch Communications in Substances of Abuse
Volume4
Issue number1
StatePublished - 1983

Fingerprint

Membranes
Brain
Osmolar Concentration
Inhibitory Concentration 50
Opiate Alkaloids
Phencyclidine Receptors
SK&F 10047
Phencyclidine
Naloxone
Trypsin
Buffers
Hot Temperature
Binding Sites
Temperature

ASJC Scopus subject areas

  • Medicine (miscellaneous)

Cite this

Partial characterization of the specific binding of 3H-N-allylnormetazocine (SKF 10047) to rat brain membranes. / Johnson, K. M.

In: Research Communications in Substances of Abuse, Vol. 4, No. 1, 1983, p. 33-47.

Research output: Contribution to journalArticle

@article{a550452889d24b65bc786804d2f27334,
title = "Partial characterization of the specific binding of 3H-N-allylnormetazocine (SKF 10047) to rat brain membranes",
abstract = "The specific binding of 3H(±)-N-allylnormetazocine (3H(±)-SKF 10047) to rat brain membranes was defined as that which could be displaced by 100μM (+)SKF 10047. This binding was found to be saturable and Scatchard analysis indicated the presence of three binding components with differing apparent K(D) and B(max) values. These sites varied considerably in their sensitivity to the effects of heat, trypsin, and ionic strength, with the low affinity site (K(D) = 245 nM) being least sensitive to elevated temperature and trypsinization and most sensitive to elevation of the ionic strength of the incubation buffer. A modest degree of selectivity could be attained for each of the three sites by varying the concentration of 3H(±)-SKF 10047, e.g. at 160 nM 3H(±)-SKF 10047 approximately 75{\%} of the total binding was calculated to be due to the low-affinity site. Using this technique it was found that both isomers of SKF 10047 were equi-effective in displacing binding from the low-affinity site, but (-)SKF 10047 was 150 times more potent than (+)SKF 10047 at the high-affinity site. Also, naloxone was very potent in inhibiting binding to the high-affinity site (IC50 = 4.7 nM), but very weak at the low-affinity site (IC50 = 100 μM). Phencyclidine (PCP) and several of its derivatives were found to have potencies similar to those of SKF 10047 in inhibiting binding to the low-affinity site. These data suggest that the low-affinity binding site for this prototypic sigma opiate may be analogous to the putative PCP receptor.",
author = "Johnson, {K. M.}",
year = "1983",
language = "English (US)",
volume = "4",
pages = "33--47",
journal = "Research Communications in Substances of Abuse",
issn = "0193-0818",
publisher = "PJD Publications Ltd",
number = "1",

}

TY - JOUR

T1 - Partial characterization of the specific binding of 3H-N-allylnormetazocine (SKF 10047) to rat brain membranes

AU - Johnson, K. M.

PY - 1983

Y1 - 1983

N2 - The specific binding of 3H(±)-N-allylnormetazocine (3H(±)-SKF 10047) to rat brain membranes was defined as that which could be displaced by 100μM (+)SKF 10047. This binding was found to be saturable and Scatchard analysis indicated the presence of three binding components with differing apparent K(D) and B(max) values. These sites varied considerably in their sensitivity to the effects of heat, trypsin, and ionic strength, with the low affinity site (K(D) = 245 nM) being least sensitive to elevated temperature and trypsinization and most sensitive to elevation of the ionic strength of the incubation buffer. A modest degree of selectivity could be attained for each of the three sites by varying the concentration of 3H(±)-SKF 10047, e.g. at 160 nM 3H(±)-SKF 10047 approximately 75% of the total binding was calculated to be due to the low-affinity site. Using this technique it was found that both isomers of SKF 10047 were equi-effective in displacing binding from the low-affinity site, but (-)SKF 10047 was 150 times more potent than (+)SKF 10047 at the high-affinity site. Also, naloxone was very potent in inhibiting binding to the high-affinity site (IC50 = 4.7 nM), but very weak at the low-affinity site (IC50 = 100 μM). Phencyclidine (PCP) and several of its derivatives were found to have potencies similar to those of SKF 10047 in inhibiting binding to the low-affinity site. These data suggest that the low-affinity binding site for this prototypic sigma opiate may be analogous to the putative PCP receptor.

AB - The specific binding of 3H(±)-N-allylnormetazocine (3H(±)-SKF 10047) to rat brain membranes was defined as that which could be displaced by 100μM (+)SKF 10047. This binding was found to be saturable and Scatchard analysis indicated the presence of three binding components with differing apparent K(D) and B(max) values. These sites varied considerably in their sensitivity to the effects of heat, trypsin, and ionic strength, with the low affinity site (K(D) = 245 nM) being least sensitive to elevated temperature and trypsinization and most sensitive to elevation of the ionic strength of the incubation buffer. A modest degree of selectivity could be attained for each of the three sites by varying the concentration of 3H(±)-SKF 10047, e.g. at 160 nM 3H(±)-SKF 10047 approximately 75% of the total binding was calculated to be due to the low-affinity site. Using this technique it was found that both isomers of SKF 10047 were equi-effective in displacing binding from the low-affinity site, but (-)SKF 10047 was 150 times more potent than (+)SKF 10047 at the high-affinity site. Also, naloxone was very potent in inhibiting binding to the high-affinity site (IC50 = 4.7 nM), but very weak at the low-affinity site (IC50 = 100 μM). Phencyclidine (PCP) and several of its derivatives were found to have potencies similar to those of SKF 10047 in inhibiting binding to the low-affinity site. These data suggest that the low-affinity binding site for this prototypic sigma opiate may be analogous to the putative PCP receptor.

UR - http://www.scopus.com/inward/record.url?scp=0020962230&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0020962230&partnerID=8YFLogxK

M3 - Article

VL - 4

SP - 33

EP - 47

JO - Research Communications in Substances of Abuse

JF - Research Communications in Substances of Abuse

SN - 0193-0818

IS - 1

ER -