Use of specific antibodies to quantitate the guanine nucleotide-binding protein Go in brain.
- 1 April 1986
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 83 (7), 2258-2262
- https://doi.org/10.1073/pnas.83.7.2258
Abstract
We immunized rabbits with purified guanine nucleotide-binding proteins (G proteins) from bovine brain and obtained an antiserum, RV3, that reacts specifically with the .alpha. subunit (39 kDa) of a G protein of unknown function, termed Go, as well as with the .beta. subunit (35 kDa) common to all G proteins. RV3 showed no crossreactivity with the .alpha. subunits of the stimulatory (Gs) or inhibitory (Gi) G proteins associated with adenylate cyclase, nor with that of the rod outer segment G protein, transducin. Immunoblots with crude and affinity-purified antiserum showed that RV3 specifically recognizes the Go .alpha. subunit and the .beta. subunit in crude brain membranes. Using RV3, we found approximately equal amounts of Go in brain membranes from frog, chicken, rat, cow, and man. Quantitative immunoblotting gave Go .alpha. subunit/.beta. subunit ratios .apprxeq. 1 in cerebral cortex, raising the possibility that free Go .alpha. subunit (unassociated with .beta. subunit) may exist in brain. The concentration of Go .alpha. subunit in cerebral cortex is about 5 times that of Gi .alpha. subunit. The results show that Go is an immunochemically distinct, highly conserved protein distributed throughout the brain, with particularly high concentrations in forebrain.This publication has 19 references indexed in Scilit:
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