Small-molecule inhibitor leads of ribosome-inactivating proteins developed using the doorstop approach.
Small-molecule inhibitor leads of ribosome-inactivating proteins developed using the doorstop approach.
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DOI：
10.1371/journal.pone.0017883
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发表时间：
2011-03-24
期刊：
影响因子：
3.7
通讯作者：
Millard CB
中科院分区：
文献类型：
--
作者：
Pang YP;Park JG;Wang S;Vummenthala A;Mishra RK;McLaughlin JE;Di R;Kahn JN;Tumer NE;Janosi L;Davis J;Millard CB
核糖体失活蛋白（RIP）是有毒的，因为它们与28 S rRNA结合并从α-八叠球菌素/蓖麻毒素环（SRL）中脱嘌呤，从而抑制蛋白质合成。志贺样毒素（Stx 1和Stx 2）是由大肠杆菌产生的，是引起食源性疾病暴发的RIP，具有显著的发病率和死亡率。蓖麻毒素是蓖麻子植物产生的，是另一种对哺乳动物致命的RIP。目前，没有美国食品和药物管理局批准的疫苗或治疗方法可以预防蓖麻毒素，志贺样毒素或其他RIP。开发有效的小分子RIP抑制剂作为治疗剂是具有挑战性的，因为RIP·SRL界面处的强静电相互作用使得药物样分子在与rRNA竞争结合RIP时无效。在本文中，我们报告了在300 nM的药物浓度下显示出高达20%的细胞保护作用以对抗蓖麻毒素或Stx 2的小分子。这些分子是使用门挡方法发现的，这是一种蛋白质多核苷酸抑制剂的新方法，它将小分子识别为门挡，以防止RIP的活性位点残基（例如，蓖麻毒素的Tyr 80或Stx 2的Tyr 77）采用活性构象，从而阻断蛋白质的功能，而不是竞争结合RIP的竞争者。这项工作为开发RIP疗法提供了有希望的线索。结果表明，门挡方法也可能适用于开发其他蛋白质·多核苷酸抑制剂作为抗病毒剂，例如痘病毒中Z-DNA结合蛋白的抑制剂。这项工作还要求对从化学筛选中获得的药物先导物进行仔细的化学和生物学表征，以避免识别不相关的化学结构，并避免筛选的化学物质与筛选测定中使用的荧光素酶报告基因之间的直接相互作用引起的干扰。
Ribosome-inactivating proteins (RIPs) are toxic because they bind to 28S rRNA and depurinate a specific adenine residue from the α-sarcin/ricin loop (SRL), thereby inhibiting protein synthesis. Shiga-like toxins (Stx1 and Stx2), produced by Escherichia coli, are RIPs that cause outbreaks of foodborne diseases with significant morbidity and mortality. Ricin, produced by the castor bean plant, is another RIP lethal to mammals. Currently, no US Food and Drug Administration-approved vaccines nor therapeutics exist to protect against ricin, Shiga-like toxins, or other RIPs. Development of effective small-molecule RIP inhibitors as therapeutics is challenging because strong electrostatic interactions at the RIP•SRL interface make drug-like molecules ineffective in competing with the rRNA for binding to RIPs. Herein, we report small molecules that show up to 20% cell protection against ricin or Stx2 at a drug concentration of 300 nM. These molecules were discovered using the doorstop approach, a new approach to protein•polynucleotide inhibitors that identifies small molecules as doorstops to prevent an active-site residue of an RIP (e.g., Tyr80 of ricin or Tyr77 of Stx2) from adopting an active conformation thereby blocking the function of the protein rather than contenders in the competition for binding to the RIP. This work offers promising leads for developing RIP therapeutics. The results suggest that the doorstop approach might also be applicable in the development of other protein•polynucleotide inhibitors as antiviral agents such as inhibitors of the Z-DNA binding proteins in poxviruses. This work also calls for careful chemical and biological characterization of drug leads obtained from chemical screens to avoid the identification of irrelevant chemical structures and to avoid the interference caused by direct interactions between the chemicals being screened and the luciferase reporter used in screening assays.
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影响因子：
1.8
作者：
Braibante, MEF;Braibante, HTS;Martins, DB
通讯作者：
Martins, DB
DOI：
10.1039/jr9530003802
发表时间：
1953-01-01
期刊：
JOURNAL OF THE CHEMICAL SOCIETY
影响因子：
--
作者：
DAVOLL, J
通讯作者：
DAVOLL, J
DOI：
10.1073/pnas.0911606106
发表时间：
2009-12-01
影响因子：
11.1
作者：
Ho, Meng-Chiao;Sturm, Matthew B.;Schramm, Vern L.
通讯作者：
Schramm, Vern L.
影响因子：
2.7
作者：
Dooley AJ;Shindo N;Taggart B;Park JG;Pang YP
通讯作者：
Pang YP
影响因子：
7.3
作者：
Heitman, Laura H.;van Veldhoven, Jacobus P. D.;IJzerman, Adriaan P.
通讯作者：
IJzerman, Adriaan P.