Levels of Gsα(short and long), Gα(olf) and Gβ(common) subunits, and calcium-sensitive adenylyl cyclase isoforms (1, 5/6, 8) in post-mortem human brain caudate and cortical membranes: comparison with rat brain membranes and potential stoichiometric relationships.
The levels of expression of Gsα(short and long), Gα(olf) and Gβ(common) subunits, and calcium-sensitive adenylyl cyclases isoforms (AC1, 5/6, and 8) in human brain cortical and caudate membranes were quantified by western blot analysis in order to establish their contribution to the patterns of AC functioning. Both areas expressed Gsα(long) (52 kDa) with values ranging from about 1400 ng/mg of membrane protein in cerebral cortex to close to 600 ng/mg of membrane protein in caudate nucleus. In contrast, Gsα(short) and Gsα(olf) were expressed separately, Gsα(short) in cortical membranes with values around 500 ng/mg of membrane protein and Gα(olf) in caudate membranes with values around 1300 ng/mg of membrane protein. Quantitative measurements of Gβ, revealed a similar expression level in cortical and caudate membranes (5444±732 versus 5511±394 ng/mg protein; p=0.966). The B(max) values of GTPγS-dependent [(3)H]-forskolin binding show the following descending order: rat striatal membranes>rat cortical membranes=human caudate membranes>human cortical membranes. Therefore, as measured immunochemically and by [(3)H]-forskolin binding, there seems to be a vast excess of Gsα subunits over catalytic units of AC. The highest levels of AC5/6 expression were detected in caudate membranes. AC8 was little expressed, and there were no significant differences in the relative values between both human brain regions. Finally, the levels of the AC1 isoform were significantly lower in caudate than in cortical membranes. It is concluded that these stoichiometric data contribute nonetheless to explain the significant differences observed in signalling capacities through the AC system in both human brain regions.
Department of Pharmacology, Faculty of Pharmacy (Vitoria-Gasteiz), University of the Basque Country (UPV/EHU), Spain.
Amaia
A
Zalduegui
Maider
M
López de Jesús
Luis F
LF
Callado
J Javier
JJ
Meana
Joan
J
Sallés
D000262:Adenylyl Cyclases / Q000378:metabolism*; D000328:Adult; D000818:Animals; D002421:Caudate Nucleus / Q000378:metabolism*; D002540:Cerebral Cortex / Q000378:metabolism*; D005260:Female; D044385:GTP-Binding Protein alpha Subunits / Q000378:metabolism; D019205:GTP-Binding Protein alpha Subunits, Gs / Q000378:metabolism; D044387:GTP-Binding Protein beta Subunits / Q000378:metabolism; D016244:Guanosine 5'-O-(3-Thiotriphosphate) / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D011485:Protein Binding / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D013045:Species Specificity
D044385:GTP-Binding Protein alpha Subunits; D044387:GTP-Binding Protein beta Subunits; D007527:Isoenzymes; D008565:Membrane Proteins; C084618:olfactory G protein subunit alpha olf; D016244:Guanosine 5'-O-(3-Thiotriphosphate); D019205:GTP-Binding Protein alpha Subunits, Gs; D000262:Adenylyl Cyclases; C106670:adenylyl cyclase 1; C121100:adenylyl cyclase 6; C118292:adenylyl cyclase 8