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Use of Monoclonal Antibodies to Study the Structural Basis of the Function of Nicotinic Acetylcholine Receptors on Electric Organ and Muscle and to Determine the Structure of Nicotinic Acetylcholine Receptors on Neurons
Annual rept. (Final) 15 Feb 1989-30 Jun 1989
SALK INST FOR BIOLOGICAL STUDIES LA JOLLA CA RECEPTOR BIOLOGY LAB
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Extensive progress has been made on studies of the structure of nicotinic acetylcholine receptors from muscles and nerves. This has provided altogether new insights into the structure of neuronal nicotinic receptors Muscle-type receptors from Torpedo electric organs were shown to be composed of subunits organized like barrels staves around a central cation channel, in the order alpha beta alpha gamma sigma. Specific amino acid sequence were mapped topographically and to various functional domains. Xenopus oocytes were found to be a good expression system for small amounts of receptor. A human cell line was shown to synthesize relatively large amounts of functional receptors composed of alpha, Beta, gamma, and sigma subunits, and to synthesize nearly equal amounts of unassembled, partially mature alpha subunits. cDNAs for alpha and sigma subunits were cloned and sequenced. Keywords Protein sequencing, Protein purification, Ligands, Monoclonal antibodies, Genes, Ion channels, Bungaro toxins.
APPROVED FOR PUBLIC RELEASE