Characterization of a purified nicotinic receptor from rat brain by using idiotypic and anti-idiotypic antibodies

Journal Article · Tue Sep 01 00:00:00 EDT 1987 · Proc. Natl. Acad. Sci. U.S.A.; (United States)

DOI:https://doi.org/10.1073/pnas.84.18.6587· OSTI ID:7241352

Abood, L G; Langone, J J; Bjercke, R; Lu, X; Banerjee, S

The availability of an anti-nicotine monoclonal antibody has made it possible to further establish the nature of the nicotine recognition proteins purified from rat brain by affinity chromatography and to provide a highly sensitive assay for determining (/sup 3/H)nicotine binding to the purified material. An enantiomeric analogue of nicotine. (-)-6-hydroxymethylnicotine, was used to prepare the affinity column. In addition, with the use of an anti-idiotypic monoclonal antibody, it was confirmed that the recognition site for nicotine resides on a protein complex composed of two components with molecular masses of 62 and 57 kDa. It was also demonstrated that the same two proteins could be purified by immunoaffinity chromatography with the use of an anti-idiotypic monoclonal antibody. With the use of the anti-nicotine antibody to measure (/sup 3/H)nicotine binding, the purified material was shown to bind 250 pmol/mg of protein. By utilizing a procedure in which the purified receptor protein was conjugated to membranes by disulfide bonds, a binding activity of 80 pmol/mg was obtained. With the availability of sterospecific monoclonal antibodies to (-)-nicotine as well as monoclonal anti-idiotypic antibodies derived when the anti-nicotine antibodies were used as immunogens, additional procedures became available for the further characterization of the purified nicotine receptor and examining its (-)-(/sup 3/H)nicotine-binding characteristics.

• Research Organization: Univ. of Rochester Medical Center, NY (USA)
• OSTI ID: 7241352
• Journal Information: Proc. Natl. Acad. Sci. U.S.A.; (United States), Journal Name: Proc. Natl. Acad. Sci. U.S.A.; (United States) Vol. 84:18; ISSN PNASA
• Country of Publication: United States
• Language: English

Related Subjects

550601* -- Medicine-- Unsealed Radionuclides in Diagnostics
62 RADIOLOGY AND NUCLEAR MEDICINE
ALKALOIDS
AMINES
ANIMALS
ANTIBODIES
AUTONOMIC NERVOUS SYSTEM AGENTS
AZINES
AZOLES
BODY
BRAIN
CELL CONSTITUENTS
CELL MEMBRANES
CENTRAL NERVOUS SYSTEM
CHROMATOGRAPHY
CONFIGURATION INTERACTION
DRUGS
ELECTROPHORESIS
HETEROCYCLIC COMPOUNDS
LABELLED COMPOUNDS
MAMMALS
MEMBRANE PROTEINS
MEMBRANES
MONOCLONAL ANTIBODIES
NERVOUS SYSTEM
NICOTINE
ORGANIC COMPOUNDS
ORGANIC NITROGEN COMPOUNDS
ORGANS
PARASYMPATHOLYTICS
PARASYMPATHOMIMETICS
PROTEINS
PYRIDINES
PYRROLES
PYRROLIDINES
RADIOCHROMATOGRAPHY
RATS
RECEPTORS
RODENTS
SEPARATION PROCESSES
TRITIUM COMPOUNDS
VERTEBRATES