STRUCTURE AND FUNCTIONS OF G-PROTEIN BETA-GAMMA SUBUNITS

G 蛋白 β-γ 亚基的结构和功能

• 批准号: 3297516
• 负责人: JOHN K NORTHUP
• 金额: $ 20.69万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1988
• 资助国家: 美国
• 起止时间: 1988-04-01 至 1992-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3297516
• 关键词: ADP ribosylation; G protein; adenosine diphosphate; adenylate cyclase; adrenal glands; affinity chromatography; alpha antiadrenergic agent; angiotensin II; chick embryo; complementary DNA; cyanogen bromide; guanosine triphosphate; laboratory rabbit; liver cells; monoclonal antibody; muscarinic receptor; muscle cells; myofibrils; pertussis toxin; phospholipase C; platelets; potassium channel; protein structure function

The structural and functional homology of the beta and gamma subunit complexes from signal transducing GTP-binding proteins (G-proteins) will be examined. The proposed research is aimed at establishing the degree to which the beta gamma complexes from several purified G-proteins and the structurally distinct forms of beta gamma proteins have the identical functions. Two functional assays will be established to characterize the beta gamma proteins: 1. The enhancement of pertussis toxin-catalyzed ADP- ribosylation of resolved alpha subunits of Gt, Gi, and Go proteins; and 2. The regulation of the relative affinities of G-protein alpha subunits for GDP and GTP. The apparently distinct form of beta gamma protein, beta-35, will be partially sequenced for protease and CNBr fragments which are electrophoretically or immunologically distinct from those of the Gt beta subunit. The structures for the immunologically distinct gamma-peptides of rabbit liver Gi, human placental Gi, and bovine Go will be determined by molecular cloning of cDNAs. The participation of specific beta gamma protein(s) in the transductions of the G- protein mediated alpha-adrenergic inhibition of human platelet adenylyl cyclase, muscarinic activation of voltage-dependent potassium channels in isolated chick myocytes, and angiotensin II activation of phospholipase C of bovine adrenal cortex and rat hepatocytes will be probed using rabbit polyclonal antisera or affinity purified fractions therefrom. The interaction of beta gamma with a low molecular weight GTP-binding protein, Gp, will be investigated, in order to determine if two distinct classes of G- proteins exist characterized differentially by beta gamma interaction. Finally, a beta gamma affinity chromatography for interacting proteins will be developed to isolate them for identification of cellular function(s). This set of studies should allow for the evaluation, quantitatively and qualitatively, of existing hypotheses concerning the cellular function of the beta gamma subunits of signal transducing G-proteins.

β和γ的结构和功能同源性 来自信号转导 GTP 结合蛋白的亚基复合物 （G 蛋白）将被检查。 拟议的研究旨在 确定 β-γ 复合物的程度 几种纯化的 G 蛋白和结构上不同的形式 β-γ蛋白具有相同的功能。 两个功能 将建立测定来表征 β 伽马 蛋白质： 1.增强百日咳毒素催化的ADP- Gt、Gi 和 Go 蛋白的已解析 α 亚基的核糖基化； 2. G蛋白α相对亲和力的调节 GDP 和 GTP 的子单位。 明显不同的 beta 形式 γ 蛋白，β-35，将进行部分蛋白酶测序 和 CNBr 片段，其是电泳或 免疫学上与 Gt β 亚基不同。 这 免疫学上不同的γ-肽的结构 兔肝 Gi、人胎盘 Gi 和牛 Go 将 通过 cDNA 的分子克隆来确定。 的参与 G-转导中的特定β-γ蛋白 蛋白质介导的人血小板α-肾上腺素能抑制 腺苷酸环化酶，电压依赖性毒蕈碱激活 离体鸡肌细胞中的钾通道和血管紧张素 II 牛肾上腺皮质和大鼠磷脂酶C的激活 将使用兔多克隆抗血清或 由其亲和纯化的级分。 贝塔的相互作用 γ 具有低分子量 GTP 结合蛋白 Gp，将 进行调查，以确定是否有两个不同类别的 G- 蛋白质的存在具有不同的β-γ特征 相互作用。 最后，β-γ亲和层析 将开发相互作用的蛋白质来分离它们 细胞功能的鉴定。 这组研究应该 允许进行定量和定性评估 关于β细胞功能的现有假设 信号转导G蛋白的γ亚基。

项目成果

Rhodopsin and the retinal G-protein distinguish among G-protein beta gamma subunit forms.

• DOI: 10.1016/s0021-9258(18)98880-0
• 发表时间: 1991-07
• 期刊: The Journal of biological chemistry
• 作者: A. Fawzi;D. Fay;E. A. Murphy;H. Tamir;J. J. Erdos-J.;J. Northup

G-protein beta gamma forms: identity of beta and diversity of gamma subunits.

G 蛋白 β γ 形式：β 的同一性和 γ 亚基的多样性。

• DOI: 10.1021/bi00230a018
• 发表时间: 1991
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Tamir,H;Fawzi,AB;Tamir,A;Evans,T;Northup,JK
• 通讯作者: Northup,JK

Unique guanine nucleotide binding properties of the human placental GTP-binding protein Gp.

人胎盘 GTP 结合蛋白 Gp 具有独特的鸟嘌呤核苷酸结合特性。

• DOI: 10.1021/bi00482a002
• 发表时间: 1990
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Tamir,A;Fawzi,AB;Northup,JK
• 通讯作者: Northup,JK

Defective guanyl nucleotide-binding protein beta gamma subunits in a forskolin-resistant mutant of the Y1 adrenocortical cell line.

Y1 肾上腺皮质细胞系的毛喉素抗性突变体中存在缺陷的鸟苷酸结合蛋白 β γ 亚基。

• DOI: 10.1073/pnas.89.19.8933
• 发表时间: 1992
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Mitchell,J;Northup,JK;Schimmer,BP
• 通讯作者: Schimmer,BP