Characterisation and partial purification of the GABA(B) receptor from the rat cerebellum using the novel antagonist [3H]CGP 62349

Autor:	M J, Keir, M J, Barakat, K K, Dev, H, Bittiger, B, Bettler, J M, Henley
Rok vydání:	1999
Předmět:	Binding Sites Detergents Transfection Benzoates Rats Kinetics Organophosphorus Compounds Receptors GABA-B Cerebellum COS Cells Centrifugation Density Gradient Animals Biotinylation Electrophoresis Polyacrylamide Gel Isonicotinic Acids GABA-B Receptor Antagonists Cells Cultured
Zdroj:	Brain research. Molecular brain research. 71(2)
ISSN:	0169-328X
Popis:	The novel GABA(B) receptor antagonist [3H]CGP 62349 binds rat cerebellar synaptosomal membranes with high affinity at a single population of sites (K(d) = 0.9 nM, B(max) = 760 fmol/mg protein). Solubilisation with 1% Triton X-100/0.5 M NaCl/10% glycerol resulted in a marked increase in [3H]CGP 62349 binding (K(d) = 0.5 nM, B(max) = 1285 fmol/mg protein). Competition of [3HCGP 35348 = CGP 36742. The GABA(A) ligand isoguvacine did not displace [3H]CGP 62349 binding. Partial purification of [3H]CGP 62349 binding sites was obtained by sucrose density centrifugation and a predominant protein in the peak binding fraction was recognised by an anti-GABA(B) receptor antibody and had a molecular weight similar to the recombinant expressed GABA(B)R1a. These results demonstrate that [3H]CGP 62349 provides a useful additional tool for further characterisation of the pharmacology and biochemistry of the native GABA(B) receptor.
Databáze:	OpenAIRE
Externí odkaz:	https://explore.openaire.eu/search/publication?articleId=pmid________::eed2cd3f7965cb99ba3182b03ca43793 https://pubmed.ncbi.nlm.nih.gov/10521582 Zobrazit plný text záznamu Full Text from ScienceDirect