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Expression, purification, and characterization of multiple, multifunctional human glucocorticoid receptor proteins

机译:多种多功能人糖皮质激素受体蛋白的表达,纯化和表征

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摘要

The glucocorticoid receptor (GR) is a nuclear receptor protein that plays a central role in glucose homeostasis, the stress response, control of the hypothalamic-pituitary-adrenal axis, and immuno-inflammatory processes via binding of the natural steroid, cortisol. GR is a well-validated drug target and continues to be an important target for new drug discovery efforts. Here, we describe a basic and simple method for Escherichia coli expression and purification of a variety of human GR proteins that contain all three of the functional domains of the protein: the activation function-1 domain, the DNA-binding domain, and the ligand-binding domain. We present characterization data to show that these purified, multifunctional GR proteins are active for ligand, coactivator, and DNA-binding. The work presented here should serve as a reference for future mechanistic, structural and drug discovery efforts that require purified, full or near full length, GR protein. (C) 2008 Elsevier Inc. All rights reserved.
机译:糖皮质激素受体(GR)是一种核受体蛋白,在葡萄糖稳态,应激反应,下丘脑-垂体-肾上腺轴的控制以及通过天然类固醇皮质醇的结合引起的免疫炎症过程中起着核心作用。 GR是经过充分验证的药物靶标,并且仍然是新药开发工作的重要靶标。在这里,我们描述了一种用于大肠杆菌表达和纯化多种人类GR蛋白的基本且简单的方法,这些GR蛋白包含该蛋白的所有三个功能域:激活功能1域,DNA结合域和配体绑定域。我们提供表征数据,以显示这些纯化的多功能GR蛋白对配体,共激活剂和DNA结合具有活性。此处介绍的工作应作为未来需要纯化,全长或接近全长GR蛋白的机理,结构和药物发现工作的参考。 (C)2008 Elsevier Inc.保留所有权利。

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