Comparison of the molecular structure of GABA/benzodiazepine receptors purified from rat and human cerebellum.
The use of improved affinity chromatographic techniques has allowed for the substantial copurification of both the benzodiazepine and the GABA receptor from brain. These preparations have been used to begin characterization of the benzodiazepine receptor at a molecular level. We have recently purified benzodiazepine receptor from human and rat cerebellum, and SDS-PAGE has revealed that both preparations consist of a major protein of 50 kDa and a minor protein of 55 kDa. These proteins are recognized by a series of monoclonal antibodies prepared against the rat benzodiazepine receptor suggesting the rat and human receptors share several common antigenic domains. Several other approaches have been employed to further investigate possible homology between the rat and human receptors. Proteolytic degradation studies have shown that the major limiting photolabeled peptide fragment generated in rat and human is similar as determined by HPLC analysis. Isoelectric focusing and SDS (two-dimensional) electrophoresis have revealed that the immunoreactive, photolabeled 50 kDa protein, and the purified receptor have identical PI values. The receptor from both human and rat are glycoproteins as determined by lectin binding studies. However, exposure of these proteins to neuraminidase fails to alter the pharmacology of the receptors indicating possible similarities in their posttranslational glycosylation. Thus, it appears that some degree of structural homology exists between the rat and human benzodiazepine receptors.
Sweetnam
P
P
Department of Psychiatry, Yale University, Connecticut Mental Health Center, New Haven 06508.
Nestler
E
E
Gallombardo
P
P
Brown
S
S
Duman
R
R
Bracha
H S
HS
Tallman
J
J
D000818:Animals; D002531:Cerebellum / Q000032:analysis*; D006026:Glycoside Hydrolases / Q000494:pharmacology; D006801:Humans; D008297:Male; D008968:Molecular Conformation; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000494:pharmacology; D051381:Rats; D011919:Rats, Inbred Strains; D011963:Receptors, GABA-A / Q000302:isolation & purification*; D013045:Species Specificity
D011963:Receptors, GABA-A; D006026:Glycoside Hydrolases; D010447:Peptide Hydrolases