Covalent Fragment Screening Identifies Rgl2 RalGEF Cysteine for Targeted Covalent Inhibition of Ral GTPase Activation.
Covalent Fragment Screening Identifies Rgl2 RalGEF Cysteine for Targeted Covalent Inhibition of Ral GTPase Activation.
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共价片段筛选用于靶向共价抑制Ral GT3活化的Ral Rgl2 RalGEF半胱氨酸。
DOI：
10.1002/cmdc.202100750
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发表时间：
2022-03-18
期刊：
影响因子：
3.4
通讯作者：
Meroueh, Samy O.
中科院分区：
文献类型：
--
作者：
Bum-Erdene, Khuchtumur;Ghozayel, Mona K.;Xu, David;Meroueh, Samy O.
关键词：
Ral GTP酶属于RAS超家族，由K-RAS直接激活。Ralgef途径是K-Ras信号转导的三条主要途径之一。按照导致K-RAS G12C治疗剂的策略，Ral GTP酶不具有半胱氨酸亲核试剂来开发共价抑制剂。然而，几种半胱氨酸氨基酸存在于鸟嘌呤交换因子的表面，可以激活Ral GTP酶，如Rgl2。在这里，我们筛选半胱氨酸电泳性片段的文库，以确定在Rgl2表面的一个半胱氨酸上形成的共价键是否可以抑制al GTP酶的激活。我们发现了几个氯乙酰胺和丙烯酰胺片段，它们抑制了Rgl2对Ral GTP酶的交换。定点突变表明，Cys-284上形成的共价键，而不是其他半胱氨酸，会抑制Rgl2激活的Ral。通过商业文库的亚结构搜索，进一步确定了Cys-284为反应位点，并确定吲哚类片段为最有希望进一步开发的系列。CyS-284位于Ral·Rgl2界面外的一个环上，该环上有几个残基与Ral GTP酶直接接触。我们的变构共价片段抑制剂为小分子共价抑制剂的开发提供了起点，以探索动物模型中的Ral GTP酶。共价片段筛选确定了在变构Cys-284处形成共价键并抑制Rgl2 Ralgef激活Ral GTP酶的HIT候选。
Ral GTPases belong to the RAS superfamily, and they are directly activated by K-RAS. The RalGEF pathway is one of the three major K-RAS signaling pathways. Ral GTPases do not possess a cysteine nucleophile to develop a covalent inhibitor following the strategy that led to a K-RAS G12C therapeutic agent. However, several cysteine amino acids exist on the surface of guanine exchange factors that activate Ral GTPases, such as Rgl2. Here, we screen a library of cysteine electrophile fragments to determine if covalent bond formation at one of the Rgl2 surface cysteines could inhibit Ral GTPase activation. We found several chloroacetamide and acrylamide fragments that inhibited Ral GTPase exchange by Rgl2. Site-directed mutagenesis showed that covalent bond formation at Cys-284, but not other cysteines, leads to inhibition of Ral activation by Rgl2. Follow-up time- and concentration-dependent studies of derivatives identified by substructure search of commercial libraries further confirmed Cys-284 as the reaction site and identified the indoline fragments as the most promising series for further development. Cys-284 is located outside of the Ral•Rgl2 interface on a loop that has several residues that come in direct contact with Ral GTPases. Our allosteric covalent fragment inhibitors provide a starting point for the development of small-molecule covalent inhibitors to probe Ral GTPases in animal models. Covalent fragment screening identified hit candidates that formed a covalent bond at allosteric Cys-284 and inhibited Rgl2 RalGEF activation of Ral GTPase.
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影响因子：
15
作者：
Kathman SG;Span I;Smith AT;Xu Z;Zhan J;Rosenzweig AC;Statsyuk AV
通讯作者：
Statsyuk AV
DOI：
10.1097/jto.0000000000000007
发表时间：
2013-12
期刊：
Journal of thoracic oncology : official publication of the International Association for the Study of Lung Cancer
影响因子：
--
作者：
Guin S;Ru Y;Wynes MW;Mishra R;Lu X;Owens C;Barn AE;Vasu VT;Hirsch FR;Kern JA;Theodorescu D
通讯作者：
Theodorescu D
DOI：
10.1073/pnas.1812963116
发表时间：
2019-02-12
影响因子：
11.1
作者：
Hillig, Roman C.;Sautier, Brice;Bader, Benjamin
通讯作者：
Bader, Benjamin
影响因子：
14.8
作者：
Cherfils, Jacqueline;Zeghouf, Mahel
通讯作者：
Zeghouf, Mahel
影响因子：
8
作者：
Cardoso, Rosa;Love, Robert;Brooun, Alexei
通讯作者：
Brooun, Alexei