Identification of hydrophobic tags for the degradation of stabilized proteins.
Identification of hydrophobic tags for the degradation of stabilized proteins.
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DOI：
10.1002/cbic.201100793
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发表时间：
2012-03-05
期刊：
影响因子：
3.2
通讯作者：
Crews, Craig M.
中科院分区：
文献类型：
--
作者：
Tae, Hyun Seop;Sundberg, Thomas B.;Neklesa, Taavi K.;Noblin, Devin J.;Gustafson, Jeffrey L.;Roth, Anke G.;Raina, Kanak;Crews, Craig M.
传统上，生物学家依靠温度敏感突变体、转录开关或 RNAi 对编码转录物的降解来调节复杂生物系统中目的蛋白 (POI) 的功能。 [1, 2] 尽管这些系统很实用，但其提供的条件控制比小分子抑制剂要少，这一限制对于长寿命和/或高丰度蛋白质来说更为严重，并且可能难以在活体动物中使用。不幸的是，尽管化学遗传筛选已成功鉴定出具有新靶点的有用“探针”化合物，但许多生物学上有趣的蛋白质（例如支架蛋白和转录因子）缺乏特异性、高亲和力的小分子配体。[3, 4]或者，一些研究小组已经开发出使用细胞可渗透的小分子配体调节蛋白质丰度的通用方法，该配体用于与 POI 基因融合的受体。[5]通常用于此目的的一组相互作用是雷帕霉素、FK506 或这些天然产物的衍生物与脯氨酰异构酶 FKB12 或哺乳动物雷帕霉素靶点 (FRB) 的 FKB12 结合结构域的结合。例如，这种方法已被用来将 POI 募集到蛋白酶体或线粒体外膜，从而分别导致其降解或错误定位。[6, 7] 然而，这些方法需要将两种融合蛋白引入细胞中。为了解决这个问题，一些实验室开发了将 POI 融合到不稳定蛋白质片段（称为降解域 (DD)）的方法，该片段在没有生物正交“稳定”配体的情况下会快速降解。[8-11] 尽管基于 DD 的系统已用于体内研究，但它们在长期实验中的实用性因注射稳定配体之间 POI 丰度的潜在波动而受到影响。[12, 13] 更多最近，神秘的 DD 的暴露已被用来降解 FKBP12-POI 融合体，以响应
Biologists have traditionally relied on temperature-sensitive mutants, transcriptional switches or degradation of coding transcripts by RNAi to regulate the function of a proteinof-interest (POI) in complex biological systems.[1, 2] Despite their utility, these systems offer less conditional control than small molecule inhibitors, a limitation that is exaggerated for long-lived and/or highly abundant proteins, and can be difficult to use in live animals. Unfortunately, although chemical genetic screening has successfully identified useful “probe” compounds with novel targets, many biologically interesting proteins (eg, scaffolding proteins and transcription factors) lack specific, high-affinity small molecule ligands.[3, 4]Alternatively, several groups have developed general methods to regulate protein abundance using cell-permeable small molecule ligands for a receptor genetically fused to a POI.[5] A set of interactions commonly used for this purpose is binding of the prolyl isomerase FKB12 or the FKB12-binding domain of mammalian target of rapamycin (FRB) by rapamycin, FK506 or derivatives of these natural products. For instance, this approach has been used to recruit a POI to the proteasome or mitochondrial outer membrane, thereby leading to its degradation or mislocalization, respectively.[6, 7] However, these methods require the introduction of two fusion proteins into cells. To address this issue, several labs have developed methods in which a POI is fused to an unstable protein fragment, termed a degradation domain (DD), that is rapidly degraded in the absence of bio-orthogonal “stabilizing” ligands.[8–11] Although DD-based systems have been used for in vivo studies, their utility for long-term experiments is compromised by the potential for fluctuations in POI abundance between injections of the stabilizing ligand.[12, 13] More recently, exposure of a cryptic DD has been used to degrade FKBP12-POI fusions in response to binding of a
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影响因子：
16
作者：
Wang Q;Liu Y;Soetandyo N;Baek K;Hegde R;Ye Y
通讯作者：
Ye Y
影响因子：
15
作者：
Erkelenz, Michael;Kuo, Chi-Hsien;Niemeyer, Christof M.
通讯作者：
Niemeyer, Christof M.
影响因子：
11.8
作者：
Robinson MS;Sahlender DA;Foster SD
通讯作者：
Foster SD
影响因子：
82.9
作者：
Banaszynski, Laura A.;Sellmyer, Mark A.;Contag, Christopher H.;Wandless, Thomas J.;Thorne, Steve H.
通讯作者：
Thorne, Steve H.
DOI：
10.1126/science.1188191
发表时间：
2010-05-14
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Dvorin JD;Martyn DC;Patel SD;Grimley JS;Collins CR;Hopp CS;Bright AT;Westenberger S;Winzeler E;Blackman MJ;Baker DA;Wandless TJ;Duraisingh MT
通讯作者：
Duraisingh MT