Identification and characterization of the 35-kDa beta subunit of guanine-nucleotide-binding proteins by an antiserum raised against transducin.
Identification and characterization of the 35-kDa beta subunit of guanine-nucleotide-binding proteins by an antiserum raised against transducin.
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通过针对转导蛋白的抗血清对鸟嘌呤核苷酸结合蛋白的 35 kDa β 亚基进行鉴定和表征。
DOI：
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
G. Schultz
中科院分区：
文献类型：
--
作者：
W. Rosenthal;D. Koesling;U. Rudolph;C. Kleuss;M. Pallast;M. Yajima;G. Schultz
抗血清针对视网膜鸟嘌呤核苷酸结合蛋白（N 蛋白）、转导蛋白，从牛视杆细胞外节纯化。重复注射抗原后获得的血清可识别所有转导蛋白亚基（α、β和γ）。测试了一种抗血清与非视网膜 N 蛋白的交叉反应性，发现它与普遍存在的 N 蛋白、Ns 和 Ni 的 β 亚基发生交叉反应，但不与它们各自的 α 和 γ 亚基发生交叉反应。抗血清还与最近发现的 N 蛋白 No 的 β 亚基发生交叉反应，该蛋白在中枢神经系统中含量很高。这些数据支持迄今为止鉴定的 N 蛋白 β 亚基的相似性。在不使用激活配体的情况下从猪大脑皮层纯化N-蛋白产生了含有No、游离βγ复合物、Ni、No的分离的α亚基的级分以及含有不同比例的两种N-蛋白的级分。通过考马斯蓝染色的 SDS 凝胶判断，制剂的纯度至少为 80%。没有获得纯Ns。在纯化过程中使用转导蛋白抗体表明，从凝胶过滤柱中洗脱的 β 亚基主要与 Ni 和 No 的 α 亚基共洗脱，但在能够将 Ns 活性重建到 Ns 缺陷细胞系（S49 循环淋巴瘤细胞）膜中的级分中几乎检测不到。这表明在中枢神经系统中，Ni和No的浓度比Ns的浓度高很多。对从猪大脑皮层纯化的 N 蛋白进行二维凝胶电泳，可分离出 35 kDa 区域的两个主要肽，它们的 pI 值不同，并通过使用抗血清将其鉴定为 β 亚基。使用来自相同组织膜的粗胆酸盐提取物代替纯化的蛋白质获得了相同的结果。不同 β 亚基的出现可以通过翻译后 N 蛋白修饰来解释。
Antisera were raised against the retinal guanine-nucleotide-binding protein (N-protein), transducin, purified from bovine rod outer segments. Sera obtained after repeated injections of antigen recognized all transducin subunits (alpha, beta and gamma). One antiserum, tested for cross-reactivity with non-retinal N-proteins, was found to cross-react with the beta subunits of the ubiquitously occurring N-proteins, Ns and Ni, but not with their respective alpha and gamma subunits. The antiserum also cross-reacted with the beta subunit of the recently identified N-protein, No, which has been found in high abundance in the central nervous system. These data support the similarity of the beta subunits of the N-proteins identified so far. Purification of N-proteins from porcine cerebral cortex without the use of activating ligands yielded fractions containing the isolated alpha subunit of No, free beta gamma complex, Ni, No and fractions containing both N-proteins in various proportions. The purity of the preparations was at least 80% as judged by Coomassie-blue-stained SDS gels. No pure Ns was obtained. Use of the transducin antibody during the course of the purification revealed that the beta subunits coeluted from a gel filtration column largely with the alpha subunits of Ni and No but were hardly detectable in fractions that were able to reconstitute Ns activity into membranes of an Ns-deficient cell line (S49 cyc- lymphoma cells). This indicates that in the central nervous system the concentrations of Ni and No are of magnitudes higher than that of Ns. Two-dimensional gel electrophoresis of N-proteins, purified from porcine cerebral cortex, resulted in the resolution of two major peptides in the 35-kDa region, which differed in their pI values and were identified as beta subunits by the use of the antiserum. Identical results were achieved using crude cholate extracts from membranes of the same tissue instead of purified proteins. The occurrence of different beta subunits may be explained by posttranslational N-protein modification.
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Ns 和 Ni，腺苷酸环化酶的刺激性和抑制性调节成分。
DOI：
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发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Codina,J;Hildebrandt,JD;Sekura,RD;Birnbaumer,M;Bryan,J;Manclark,CR;Iyengar,R;Birnbaumer,L
通讯作者：
Birnbaumer,L
从牛中枢神经系统中分离出的 GTP 结合蛋白家族内部的关系。
DOI：
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发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Roof,DJ;Applebury,ML;Sternweis,PC
通讯作者：
Sternweis,PC
腺苷酸环化酶刺激调节成分的亚基。
DOI：
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发表时间：
1983
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Northup,JK;Smigel,MD;Sternweis,PC;Gilman,AG
通讯作者：
Gilman,AG
腺苷酸环化酶的抑制性鸟嘌呤核苷酸结合调节成分。
DOI：
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发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Katada,T;Bokoch,GM;Northup,JK;Ui,M;Gilman,AG
通讯作者：
Gilman,AG
DOI：
--
发表时间：
1985-09
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
R. Huff;J. Axton;E. Neer
通讯作者：
R. Huff;J. Axton;E. Neer