| Expression, Purification and Crystallisation of the Adenosine A2A Receptor Bound to an Engineered Mini G Protein
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Author:
Date:
2017-04-20
[Abstract] G protein-coupled receptors (GPCRs) promote cytoplasmic signalling by activating heterotrimeric G proteins in response to extracellular stimuli such as light, hormones and nucleosides. Structure determination of GPCR–G protein complexes is central to understanding the precise mechanism of signal transduction. However, these complexes are challenging targets for structural studies due to their conformationally dynamic and inherently transient nature. We recently developed an engineered G protein, mini-Gs, which addressed these problems and allowed the formation of a stable GPCR–G protein complex. Mini-Gs facilitated the structure determination of the human adenosine A2A receptor (A2AR) in its G protein-bound conformation at 3.4 Å resolution. ...
[摘要] G蛋白偶联受体（GPCR）通过激活异源三聚体G蛋白来响应细胞外刺激如光，激素和核苷来促进细胞质信号传导。 GPCR-G蛋白复合物的结构测定对于了解信号转导的精确机制至关重要。然而，由于它们的构象动态和固有的短暂性质，这些复合物是结构研究的具有挑战性的目标。我们最近开发了一种工程化的G蛋白，微型G ，解决了这些问题，并允许形成稳定的GPCR-G蛋白复合物。 Mini-G 促进了人腺苷A 2A受体（A 2A 2A）在其G蛋白结合构象中的结构测定，在3.4 Å分辨率。在这里，我们描述了A 2A R R的表达和纯化的一步一步的方案，并且A 2AA-R-mini-G'子>复杂。
背景 我们最近开发了一种工程化的最小G蛋白，迷你G（Carpenter和Tate，2016），其促进了人腺苷A 2A受体的结构测定（A <其活性状态（carpenter等人，2016）。 mini-g="">充分稳定A 2A R的活性构象，以允许络合物在洗涤剂辛硫基葡糖苷中通过蒸气扩散结晶。在这里，我们描述了一种用于表达和纯化Aβ2A ...
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| In vitro Studies: Inhibition of Nevirapine Metabolism by Nortriptyline in Hepatic Microsomes
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Author:
Date:
2015-10-05
[Abstract] One of the most prevalent and interfering psychosocial comorbidities of HIV infection is clinical depression (22 to 45%). For this reason, a study of a possible interaction between the nonnucleoside reverse transcriptase inhibitor nevirapine (NVP) and the tricyclic antidepressant nortriptyline (NT) was carried out. In vitro studies with rat and human hepatic microsomes showed a marked inhibition of NVP metabolism by NT being more intense in rat than in human. The extrapolation of these results to humans suggests increased NVP side effects when both drugs are coadministered, but additional in vivo human studies are required to evaluate the clinical implication of this interaction.
This protocol describes a technique for detecting and measuring the inhibition of ...
[摘要] HIV感染的最普遍和干扰的心理社会共病是临床抑郁症（22％至45％）。 为此，进行了非核苷逆转录酶抑制剂奈韦拉平（NVP）和三环抗抑郁药去甲替林（NT）之间可能的相互作用的研究。 使用大鼠和人肝微粒体的体外研究显示在大鼠中NT比在人中更强烈的NVP代谢的显着抑制。 将这些结果外推到人表明当两种药物共同给药时，NVP副作用增加，但是需要另外的体内人体研究来评价这种相互作用的临床意义。
该方案描述了 检测和测量肝微粒体中去甲替林对奈韦拉平代谢的抑制的技术。
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