Demonstration of sub-nanomolar affinity of bryostatin 1 for the phorbol ester receptor in rat brain

David J. de Vries, Cherry L. Herald, George Pettit, Peter M. Blumberg

Research output: Contribution to journalArticle

66 Citations (Scopus)

Abstract

The effect of bryostatin 1 on [26-3H]phorbol 12,13-dibutyrate ([3H]PDBu) binding to a washed paniculate preparation from rat brain was examined. Bryostatin 1 inhibited phorbol ester binding at concentrations considerably lower than previously reported. As would be expected for a ligand of high affinity, the apparent displacing potency of bryostatin 1 was dependent on the concentration of tissue/binding sites included in the assay. Decreasing the concentration of [3H]PDBu binding sites to the picomolar detection limit resulted in apparent bryostatin displacing potencies in the picomolar range with these values representing an upper estimate of the true affinity. When included in saturation studies with [3H]PDBu, bryostatin 1 displayed mixed competitive/non-competitive inhibition. Using either repetitive washing or dialysis of the membrane preparation, it was not possible to reverse the inhibition produced by bryostatin 1. The greater affinity of bryostatin 1 compared to other classes of agents that act directly on protein kinase C and the stability of its association may contribute to the unique biological properties of the bryostatins.

Original languageEnglish (US)
Pages (from-to)4069-4073
Number of pages5
JournalBiochemical Pharmacology
Volume37
Issue number21
DOIs
StatePublished - Nov 1 1988

Fingerprint

Rats
Brain
Demonstrations
Bryostatins
Binding Sites
Phorbol 12,13-Dibutyrate
Dialysis
Phorbol Esters
Washing
Protein Kinase C
Limit of Detection
phorbol ester receptor
bryostatin 1
Assays
Association reactions
Tissue
Ligands
Membranes

ASJC Scopus subject areas

  • Pharmacology

Cite this

Demonstration of sub-nanomolar affinity of bryostatin 1 for the phorbol ester receptor in rat brain. / de Vries, David J.; Herald, Cherry L.; Pettit, George; Blumberg, Peter M.

In: Biochemical Pharmacology, Vol. 37, No. 21, 01.11.1988, p. 4069-4073.

Research output: Contribution to journalArticle

de Vries, David J. ; Herald, Cherry L. ; Pettit, George ; Blumberg, Peter M. / Demonstration of sub-nanomolar affinity of bryostatin 1 for the phorbol ester receptor in rat brain. In: Biochemical Pharmacology. 1988 ; Vol. 37, No. 21. pp. 4069-4073.
@article{38aee5cda4814923a34bf0091188d8a9,
title = "Demonstration of sub-nanomolar affinity of bryostatin 1 for the phorbol ester receptor in rat brain",
abstract = "The effect of bryostatin 1 on [26-3H]phorbol 12,13-dibutyrate ([3H]PDBu) binding to a washed paniculate preparation from rat brain was examined. Bryostatin 1 inhibited phorbol ester binding at concentrations considerably lower than previously reported. As would be expected for a ligand of high affinity, the apparent displacing potency of bryostatin 1 was dependent on the concentration of tissue/binding sites included in the assay. Decreasing the concentration of [3H]PDBu binding sites to the picomolar detection limit resulted in apparent bryostatin displacing potencies in the picomolar range with these values representing an upper estimate of the true affinity. When included in saturation studies with [3H]PDBu, bryostatin 1 displayed mixed competitive/non-competitive inhibition. Using either repetitive washing or dialysis of the membrane preparation, it was not possible to reverse the inhibition produced by bryostatin 1. The greater affinity of bryostatin 1 compared to other classes of agents that act directly on protein kinase C and the stability of its association may contribute to the unique biological properties of the bryostatins.",
author = "{de Vries}, {David J.} and Herald, {Cherry L.} and George Pettit and Blumberg, {Peter M.}",
year = "1988",
month = "11",
day = "1",
doi = "10.1016/0006-2952(88)90097-4",
language = "English (US)",
volume = "37",
pages = "4069--4073",
journal = "Biochemical Pharmacology",
issn = "0006-2952",
publisher = "Elsevier Inc.",
number = "21",

}

TY - JOUR

T1 - Demonstration of sub-nanomolar affinity of bryostatin 1 for the phorbol ester receptor in rat brain

AU - de Vries, David J.

AU - Herald, Cherry L.

AU - Pettit, George

AU - Blumberg, Peter M.

PY - 1988/11/1

Y1 - 1988/11/1

N2 - The effect of bryostatin 1 on [26-3H]phorbol 12,13-dibutyrate ([3H]PDBu) binding to a washed paniculate preparation from rat brain was examined. Bryostatin 1 inhibited phorbol ester binding at concentrations considerably lower than previously reported. As would be expected for a ligand of high affinity, the apparent displacing potency of bryostatin 1 was dependent on the concentration of tissue/binding sites included in the assay. Decreasing the concentration of [3H]PDBu binding sites to the picomolar detection limit resulted in apparent bryostatin displacing potencies in the picomolar range with these values representing an upper estimate of the true affinity. When included in saturation studies with [3H]PDBu, bryostatin 1 displayed mixed competitive/non-competitive inhibition. Using either repetitive washing or dialysis of the membrane preparation, it was not possible to reverse the inhibition produced by bryostatin 1. The greater affinity of bryostatin 1 compared to other classes of agents that act directly on protein kinase C and the stability of its association may contribute to the unique biological properties of the bryostatins.

AB - The effect of bryostatin 1 on [26-3H]phorbol 12,13-dibutyrate ([3H]PDBu) binding to a washed paniculate preparation from rat brain was examined. Bryostatin 1 inhibited phorbol ester binding at concentrations considerably lower than previously reported. As would be expected for a ligand of high affinity, the apparent displacing potency of bryostatin 1 was dependent on the concentration of tissue/binding sites included in the assay. Decreasing the concentration of [3H]PDBu binding sites to the picomolar detection limit resulted in apparent bryostatin displacing potencies in the picomolar range with these values representing an upper estimate of the true affinity. When included in saturation studies with [3H]PDBu, bryostatin 1 displayed mixed competitive/non-competitive inhibition. Using either repetitive washing or dialysis of the membrane preparation, it was not possible to reverse the inhibition produced by bryostatin 1. The greater affinity of bryostatin 1 compared to other classes of agents that act directly on protein kinase C and the stability of its association may contribute to the unique biological properties of the bryostatins.

UR - http://www.scopus.com/inward/record.url?scp=0023738122&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023738122&partnerID=8YFLogxK

U2 - 10.1016/0006-2952(88)90097-4

DO - 10.1016/0006-2952(88)90097-4

M3 - Article

C2 - 3190746

AN - SCOPUS:0023738122

VL - 37

SP - 4069

EP - 4073

JO - Biochemical Pharmacology

JF - Biochemical Pharmacology

SN - 0006-2952

IS - 21

ER -