Optimizing the protein switch: Altering nuclear import and export signals, and ligand binding domaine
Optimizing the protein switch: Altering nuclear import and export signals, and ligand binding domaine
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DOI：
10.1016/j.jconrel.2007.04.017
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发表时间：
2007-07-31
影响因子：
10.8
通讯作者：
Lim, Carol S.
中科院分区：
文献类型：
--
作者：
Kakar, Mudit;Davis, James R.;Lim, Carol S.
本研究优化了配体调控的定位可控蛋白构建体。由经典的核输出信号（HIV-rev、MAPKK或孕酮受体）与SV 40 T抗原型核输入信号组合制备几种构建体。还测试了不同配体结合结构域（来自糖皮质激素受体或孕酮受体的LBD）赋予对蛋白质定位的控制的能力。本研究的目的是创建构建体，其在没有配体的情况下是细胞质的，在配体的存在下是核的，并且还调节在配体诱导下易位到核的蛋白质的量。输入和输出信号强度之间的平衡对于蛋白质的整体定位至关重要。进入细胞核的蛋白质的量也受到配体剂量（10-100 nM）的影响。然而，整体的进口特性是由本地化信号的强度和所使用的LBD的固有本地化属性。这项研究确定了存在于特定隔室中的蛋白质的量可以通过使用各种强度的定位信号来调节。这些优化的定位可控蛋白质构建体可用于校正由于蛋白质的异常定位引起的疾病。(c)2007 Elsevier B. V.保留所有权利。
Ligand regulated localization controllable protein constructs were optimized in this study. Several constructs were made from a classical nuclear export signal (HIV-rev, MAPKK, or progesterone receptor) in combination with a SV40 T-antigen type nuclear import signal. Different ligand binding domains (LBDs from glucocorticoid receptor or progesterone receptor) were also tested for their ability to impart control over localization of proteins. This study was designed to create constructs which are cytoplasmic in the absence of ligand and nuclear in the presence of ligand, and also to regulate the amount of protein translocating to the nucleus on ligand induction. The balance between the strengths of import and export signals was critical for overall localization of proteins. The amount of protein entering the nucleus was also affected by the dose of ligand (10-100 nM). However, the overall import characteristics were determined by the strengths of localization signals and the inherent localization properties of the LBD used. This study established that the amount of protein present in a particular compartment can be regulated by the use of localization signals of various strengths. These optimized localization controllable protein constructs can be used to correct for diseases due to aberrant localization of proteins. (c) 2007 Elsevier B.V. All rights reserved.