In Vitro Ubiquitination Platform Identifies Methyl Ellipticiniums as Ubiquitin Ligase Inhibitors.
In Vitro Ubiquitination Platform Identifies Methyl Ellipticiniums as Ubiquitin Ligase Inhibitors.
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体外泛素化平台-作为泛素连接酶抑制剂。
DOI：
10.1177/24725552211000675
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发表时间：
2021-08
期刊：
影响因子：
3.1
通讯作者：
O'Keefe, Barry R.
中科院分区：
文献类型：
--
作者：
Wilson, Brice A. P.;Voeller, Donna;Smith, Emily A.;Wamiru, Antony;Goncharova, Ekaterina, I;Liu, Gang;Lipkowitz, Stanley;O'Keefe, Barry R.
小蛋白泛素转移到靶蛋白是一个复杂的协调过程，称为泛素化，导致蛋白质功能或稳定性的调节。泛素化的适当调节是必不可少的，而这一过程的失调与几种人类疾病有关。作为重要疾病相关途径中的中心信号传导节点的泛素化级联的一个实例是CBLB [来自Cas NS-1鼠逆转录病毒的病毒致癌基因Casitas B谱系淋巴瘤（CBL）的人类同源物]，其是一种RING指泛素连接酶（E3），其底物包括许多重要的细胞信号传导激酶。这些包括在T细胞受体和共刺激途径中起作用的免疫功能中重要的激酶、天然杀伤（NK）细胞中的Tyro/Axl/MerTK（TAM）受体家族以及生长因子受体激酶如表皮生长因子受体（EGFR）。CBLB的丧失已显示出增加先天性和适应性抗肿瘤免疫。这表明CBLB E3活性的小分子调节可以增强患者的抗肿瘤免疫力。为了探索E3的酶抑制可能导致疾病相关信号传导途径的调节的假设，我们建立了用于抑制CBLB活性的> 70，000个化学实体的高通量筛选。虽然选择CBLB作为抑制剂发现的原理验证靶点，但我们证明我们的测定可推广到监测其他泛素连接酶的活性。我们用CBLB活性的另外的基于细胞的模型进一步扩展了我们观察到的体外抑制。从这些研究中，我们证明了一类天然产物为基础的生物碱，被称为甲基ellipticiniums（ME），是能够抑制细胞内泛素连接酶。
The transfer of the small protein ubiquitin to a target protein is an intricately orchestrated process called ubiquitination that results in modulation of protein function or stability. Proper regulation of ubiquitination is essential, and dysregulation of this process is implicated in several human diseases. An example of a ubiquitination cascade that is a central signaling node in important disease-associated pathways is that of CBLB [a human homolog of a viral oncogene Casitas B-lineage lymphoma (CBL) from the Cas NS-1 murine retrovirus], a RING finger ubiquitin ligase (E3) whose substrates include a number of important cell-signaling kinases. These include kinases important in immune function that act in the T cell receptor and costimulatory pathways, the Tyro/Axl/MerTK (TAM) receptor family in natural killer (NK) cells, as well as growth factor receptor kinases like epidermal growth factor receptor (EGFR). Loss of CBLB has been shown to increase innate and adaptive antitumor immunity. This suggests that small-molecule modulation of CBLB E3 activity could enhance antitumor immunity in patients. To explore the hypothesis that enzymatic inhibition of E3s may result in modulation of disease-related signaling pathways, we established a high-throughput screen of >70,000 chemical entities for inhibition of CBLB activity. Although CBLB was chosen as a proof-of-principle target for inhibitor discovery, we demonstrate that our assay is generalizable to monitoring the activity of other ubiquitin ligases. We further extended our observed in vitro inhibition with additional cell-based models of CBLB activity. From these studies, we demonstrate that a class of natural product–based alkaloids, known as methyl ellipticiniums (MEs), is capable of inhibiting ubiquitin ligases intracellularly.
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影响因子：
7.3
作者：
Kumar, Eric A.;Yuan, Ziyan;Palermo, Nicholas Y.;Dong, Lin;Ahmad, Gulzar;Lokesh, G. L.;Kolar, Carol;Kizhake, Smitha;Borgstahl, Gloria E. O.;Band, Hamid;Natarajan, Amarnath
通讯作者：
Natarajan, Amarnath
影响因子：
4.6
作者：
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通讯作者：
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影响因子：
11.2
作者：
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通讯作者：
Lipkowitz S
影响因子：
7.3
作者：
ACTON, EM;NARAYANAN, VL;BOYD, MR
通讯作者：
BOYD, MR
影响因子：
4.8
作者：
Ettenberg, SA;Magnifico, A;Lipkowitz, S
通讯作者：
Lipkowitz, S