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Title: Binding of (/sup 3/H)forskolin to solubilized preparations of adenylate cyclase

Abstract

The binding of (/sup 3/H)forskolin to proteins solubilized from bovine brain membranes was studied by precipitating proteins with polyethylene glycol and separating (/sup 3/H)forskolin bound to protein from free (/sup 3/H)forskolin by rapid filtration. The K/sub d/ for (/sup 3/H)forskolin binding to solubilized proteins was 14 nM which was similar to that for (/sup 3/H)forskolin binding sites in membranes from rat brain and human platelets. Forskolin analogs competed for (/sup 3/H)forskolin binding sites with the same rank potency in both brain membranes and in proteins solubilized from brain membranes. (/sup 3/H)forskolin bound to proteins solubilized from membranes with a Bmax of 38 fmolmg protein which increased to 94 fmolmg protein when GppNHp was included in the binding assay. In contrast, GppNHp had no effect on (/sup 3/H)forskolin binding to proteins solubilized from membranes preactivated with GppNHp. Solubilized adenylate cyclase from non-preactivated membranes had a basal activity of 130 pmolmgmin which was increased 7-fold by GppNHp. In contrast, adenylate cyclase from preactivated membranes had a basal activity of 850 pmolmgmin which was not stimulated by GppNHp or forskolin

Authors:
;
Publication Date:
Research Org.:
Food and Drug Administration, Bethesda, MD (USA)
OSTI Identifier:
5281705
Resource Type:
Journal Article
Journal Name:
Life Sci.; (United States)
Additional Journal Information:
Journal Volume: 42:14
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CYCLASES; ENZYME ACTIVITY; PROTEINS; BIOCHEMICAL REACTION KINETICS; RECEPTORS; BLOOD PLATELETS; BRAIN; CELL MEMBRANES; MAN; RATS; TRACER TECHNIQUES; TRITIUM COMPOUNDS; ANIMALS; BIOLOGICAL MATERIALS; BLOOD; BLOOD CELLS; BODY; BODY FLUIDS; CELL CONSTITUENTS; CENTRAL NERVOUS SYSTEM; ENZYMES; ISOTOPE APPLICATIONS; KINETICS; LABELLED COMPOUNDS; LYASES; MAMMALS; MATERIALS; MEMBRANE PROTEINS; MEMBRANES; NERVOUS SYSTEM; ORGANIC COMPOUNDS; ORGANS; PRIMATES; REACTION KINETICS; RODENTS; VERTEBRATES; 550201* - Biochemistry- Tracer Techniques

Citation Formats

Nelson, C.A., and Seamon, K.B. Binding of (/sup 3/H)forskolin to solubilized preparations of adenylate cyclase. United States: N. p., 1988. Web. doi:10.1016/0024-3205(88)90167-1.
Nelson, C.A., & Seamon, K.B. Binding of (/sup 3/H)forskolin to solubilized preparations of adenylate cyclase. United States. doi:10.1016/0024-3205(88)90167-1.
Nelson, C.A., and Seamon, K.B. Fri . "Binding of (/sup 3/H)forskolin to solubilized preparations of adenylate cyclase". United States. doi:10.1016/0024-3205(88)90167-1.
@article{osti_5281705,
title = {Binding of (/sup 3/H)forskolin to solubilized preparations of adenylate cyclase},
author = {Nelson, C.A. and Seamon, K.B.},
abstractNote = {The binding of (/sup 3/H)forskolin to proteins solubilized from bovine brain membranes was studied by precipitating proteins with polyethylene glycol and separating (/sup 3/H)forskolin bound to protein from free (/sup 3/H)forskolin by rapid filtration. The K/sub d/ for (/sup 3/H)forskolin binding to solubilized proteins was 14 nM which was similar to that for (/sup 3/H)forskolin binding sites in membranes from rat brain and human platelets. Forskolin analogs competed for (/sup 3/H)forskolin binding sites with the same rank potency in both brain membranes and in proteins solubilized from brain membranes. (/sup 3/H)forskolin bound to proteins solubilized from membranes with a Bmax of 38 fmolmg protein which increased to 94 fmolmg protein when GppNHp was included in the binding assay. In contrast, GppNHp had no effect on (/sup 3/H)forskolin binding to proteins solubilized from membranes preactivated with GppNHp. Solubilized adenylate cyclase from non-preactivated membranes had a basal activity of 130 pmolmgmin which was increased 7-fold by GppNHp. In contrast, adenylate cyclase from preactivated membranes had a basal activity of 850 pmolmgmin which was not stimulated by GppNHp or forskolin},
doi = {10.1016/0024-3205(88)90167-1},
journal = {Life Sci.; (United States)},
number = ,
volume = 42:14,
place = {United States},
year = {1988},
month = {1}
}