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The Marshall W. Nirenberg Papers

Laboratory project: "Muscarinic Acetylcholine Receptors of Cultured Cell Lines" pdf (120,028 Bytes) transcript of pdf
Laboratory project: "Muscarinic Acetylcholine Receptors of Cultured Cell Lines"
Studies on muscarinic acetylcholine receptors focus both on ligand-binding and on defining the physical properties of muscarinic receptors.
Item is a photocopy.
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2 (120,028 Bytes)
1979-09 (September 1979)
Nirenberg, Marshall W.
Kline, William L.
Hurko, Orest
National Heart, Lung, and Blood Institute. Laboratory of Biochemical Genetics
This item is in the public domain. It may be used without permission.
Exhibit Category:
From Neuroblastoma to Homeobox Genes, 1976-1992
Metadata Record Annual Report of the Laboratory of Biochemical Genetics, October 1, 1978 - September 30, 1979 (September 1979)
Box Number: 6
Folder Number: 22
Unique Identifier:
Accession Number:
Document Type:
Physical Condition:
Series: Series III: Laboratory Administration, [1959]-1993
SubSeries: Annual Reports, 1960-1993
Folder: Annual Reports, 1960-1988
Project Number: Z01 HL 000012-05 LBG
Period Covered: October 1, 1978 - September 30, 1979
Title of Project: Muscarinic Acetylcholine Receptors of Cultured Cell Lines
Names, Laboratory and Institute Affiliations, and Titles of Principal Investigations and All Other Professional Personnel Engaged on the Project:
PI: Marshall Nirenberg, Chief, LBG, LBG NHLBI
Other: William L. Kline, Guest Worker, LBG NHLBI
Orest Hurko, Staff Associate, LBG NHLBI
Cooperating Units (if any): None
Lab/Branch: Laboratory of Biochemical Genetics
Section: Section on Molecular Biology
Institute and Location: NHLBI, NIH, Bethesda, MD 20205
Total Man Years: 0.25
Professional: 0.25
Other: 0
Summary of Work: Studies on: muscarinic acetylcholine receptors focus both on ligand-binding and on defining the physical properties of muscarinic receptors.
Project Description:
Major Findings: [3H]-Quinuclidinyl-benzilate (QNB) was used to study muscarinic acetylcholine receptors in NG108-15 membrane preparations. The apparent dissociation constant of [3H]-QNB is 1 x 10^(-10) M; the average NG108-15 cell possesses 30,000 specific sites for [3H]-QNB. Activation of the receptors with acetylcholine or carbachol results in cell depolarization, a small increase in cellular cGMP, and inhibition of adenylate cyclase. Cell depolarization and rise in cGMP levels desensitize in 30 sec; whereas, the inhibition of adenylate cyclase does not desensitize. Scatchard analysis revealed only one homogeneous class of [3H]-QNB binding sites; however biphasic rates of [3H]-QNB association with and dissociation from receptors were found. Evidence was obtained for the formation of a dissociable [[3H]-QNB Receptor] complex which then is converted to a form which dissociates only slowly. Hill coefficients of approximately 1.0 were found for receptor antagonists and approximately 0.5 for receptor activators. A sequential series of reactions were proposed to account for these observations and for the various states of the muscarinic acetylcholine receptor that were detected.
1. Burgermeister, W., Kline, W.L., Nirenberg, M., and Witkop, B., Mol. Pharm. 14, 751-767 (1978).
2. Hurko, O. Specific [3H]-Quinuclidinyl Benzilate binding activity in digitonin-solubilized preparations for bovine brain. Arch. Biochem. and Biophys. 190, 434-445 (1978).
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