Name: quantifying agonist activity at g protein-coupled receptors
Uploaded: 2011-12-26T12:00:00
Description: Watch this Scientific Journal Video about Quantifying Agonist Activity at G Protein-coupled Receptors at JoVE.com

这项工作是由国家卫生部批准通用汽车69829研究院的支持。

1。测量激动剂浓度反应曲线：没有构活动<ol><li>相对激动剂K B值（RA I）估计，至少有两个激动剂浓度反应曲线是必需的。任何的G蛋白偶联受体的功能反应在体外检测可测激动剂浓度可以控制和单一类型的受体介导的响应。合适的细胞检测包括测量营地 4,7 和 8,9 inositolphosphate积累在细胞株表达了重组受体。整个组织检测的例子包括 10?...

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	<li>Stephenson, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+modification+of+receptor+theory.">A modification of receptor theory.</a> British Journal of Pharmacology. 11, 379-393 (1956).</li><li>Furchgott, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+b-haloalkylamines+in+the+differentiation+of+receptors+and+in+the+determination+of+dissociation+constants+of+receptor-agonist+complexes.">The use of b-haloalkylamines in the differentiation of receptors and in the determination of dissociation constants of receptor-agonist complexes.</a> Advances in Drug Research. 3, 21-55 (1966).</li><li>Ehlert, F. J., Suga, H., Griffin, M. 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由于我们估计RA ​​I（相对K B值）的方法只需要激动剂浓度反应曲线的测量，我们的分析可以做到随时测量这些曲线。 如果一天的到，每天的实验准备工作（例如，细胞或组织）的反应中的变化是大的，每个浓度-反应曲线的响应测量可以被归的“热门”的标准激动剂估计为每一天的实验。 顶部和日志欧共体50的估计，预计...

quantifying agonist activity at g protein-coupled receptors

当激动剂激活G蛋白偶联受体（GPCRs）的人口，就会引发一个信号通路在细胞或组织的反应达到高潮。这个过程可以在一个单一的受体，受体的人口，或下游的反应程度进行分析。在这里，我们描述如何分析下游的反应，以获得激动剂亲和力常数为单一受体的活跃状态的估计。 受体表现为量子开关，有效和无效状态（图1）之间的交替。激活状态的交互与特定的G蛋白或其他信号伙伴。在配位体的情况下，非国家占主导地位。激动剂约束力增加的可能性，受体将进入活跃状态的开关，因为其亲和常数为活跃状态（K B）比为无效状态（K一）要大得多。求和随机输出的人口中的所有受体产生一个恒定的水平在时间上的受体激活。引出半最大受体激活受体激动剂浓度的倒数相当于观测到的亲和常数（K OBS），并处于活动状态的激动剂-受体复合物的一小部分是定义为疗效（ε） （图2）。 已经开发了分析，对GPCRs下游反应的方法， 使估算的K OB和相对有效性激动剂1,2。在这份报告中，我们将展示如何修改此分析来估计激动剂K B值相对另一激动剂。我们的分析，展示构活动，显示如何估计 K B的M -1的绝对单位。 我们分析激动剂浓度反应曲线3,4的方法包括全球非线性回归使用的运作模式5。我们描述一个程序使用的应用软件，棱镜（GraphPad软件公司，加利福尼亚州圣迭戈）。分析得出K产品的OB和疗效（τ）成正比的参数估计。 τKOBS之一激动剂除以另一个，估计是一个相对尺度K B（RA I）6。任何参展构活动的受体，它是可能的估计参数免费受体复合物的疗效（τSYS）成正比。在这种情况下，K B激动剂的价值相当于τKOBS /τ 系统 3 。 我们的方法是有用的，确定为受体亚型激动剂的选择性和量化通过不同的G蛋白受体激动剂信令。

Watch this Scientific Journal Video about Quantifying Agonist Activity at G Protein-coupled Receptors at JoVE.com

Quantifying Agonist Activity at G Protein-coupled Receptors

一个活跃状态激动剂的亲和力常数估算方法（<em&gt; K<sub&gt; B</sub</emG蛋白偶联受体）的描述。分析提供了绝对或相对的措施<em&gt; K<sub&gt; B</sub</em&gt;根据构受体激活是否是可测量的。我们的方法适用于各种反应受体激活下游。

量化在G蛋白偶联受体激动剂活性

When an agonist activates a population of G protein-coupled receptors (GPCRs), it elicits a signaling pathway that culminates in the response of the cell ...

Research

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多巴胺的释放突触前货箱码头在个别的可视化与FFNs

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生物学

In vivo Quantification of G Protein Coupled Receptor Interactions using Spectrally Resolved Two-photon Microscopy

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在体内 G蛋白偶联受体相互作用光谱分辨的双光子显微镜定量

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成像摹蛋白偶联受体（GPCR）介导的信号，控制趋化粘菌</em

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G蛋白偶联受体的基于荧光的钙动员含量表征

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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy

通过全内反射荧光显微镜可视化G蛋白偶联受体的单事件分辨率网格蛋白介导的内吞作用

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关联特定基因的DNA甲基化与表达的变化和星形胶质细胞的转录活性<em&gt; KCNJ10</em&gt;（Kir4.1）

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配体激活的G蛋白偶联受体内在化的检测通过共聚焦显微镜

Confocal laser scanning microscopy (CLSM) is an optical imaging technique for high-contrast imaging. It is a powerful approach to visualize fluorescent ...