USE OF SPECIFIC ANTIBODIES TO QUANTITATE THE GUANINE NUCLEOTIDE-BINDING PROTEIN-G0 IN BRAIN
USE OF SPECIFIC ANTIBODIES TO QUANTITATE THE GUANINE NUCLEOTIDE-BINDING PROTEIN-G0 IN BRAIN
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DOI：
10.1073/pnas.83.7.2258
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发表时间：
1986-04-01
影响因子：
11.1
通讯作者：
SPIEGEL, A
中科院分区：
文献类型：
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作者：
GIERSCHIK, P;MILLIGAN, G;SPIEGEL, A
我们用来自牛脑的纯化的鸟嘌呤核苷酸结合蛋白（G蛋白）免疫兔，并获得了与α-鸟嘌呤核苷酸结合蛋白特异性反应的抗血清RV 3。功能未知的G蛋白（称为Go）的β亚基（39 kDa），以及与β亚基（39 kDa）的同源性。亚基（35 kDa）是所有G蛋白共有的。RV 3显示与α-SMA没有交叉反应性。与腺苷酸环化酶相关的刺激性（Gs）或抑制性（Gi）G蛋白的亚基，也不与视杆细胞外节G蛋白、转导素相关。用粗制和亲和纯化的抗血清进行的免疫印迹显示RV 3特异性识别Go α。亚基和β亚基。亚单位的粗脑细胞膜。使用RV 3，我们在来自青蛙、鸡、大鼠、牛和人的脑膜中发现了大约等量的Go。亚基/β亚基比率1在大脑皮层，提高了可能性，自由围棋α。亚基（与β不相关）亚基）可能存在于脑中。Go α的浓度大脑皮层中的Gi α亚基是Gi α亚基的约5倍。亚单位结果表明，Go是一种免疫化学上独特的高度保守的蛋白质，分布在整个大脑中，在前脑中的浓度特别高。
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.