Department of Microbiology, Immunology, and Molecular Genetics, University of California, Los Angeles, CA 90095.
Proc Natl Acad Sci U S A. 2013 Dec 10;110(50):E4904-12. doi: 10.1073/pnas.1302334110. Epub 2013 Nov 4.
Pathogenic microorganisms and toxins have evolved a variety of mechanisms to gain access to the host-cell cytosol and thereby exert virulent effects upon the host. One common mechanism of cellular entry requires trafficking to an acidified endosome, which promotes translocation across the host membrane. To identify small-molecule inhibitors that block this process, a library of 30,000 small molecules was screened for inhibitors of anthrax lethal toxin. Here we report that 4-bromobenzaldehyde N-(2,6-dimethylphenyl)semicarbazone, the most active compound identified in the screen, inhibits intoxication by lethal toxin and blocks the entry of multiple other acid-dependent bacterial toxins and viruses into mammalian cells. This compound, which we named EGA, also delays lysosomal targeting and degradation of the EGF receptor, indicating that it targets host-membrane trafficking. In contrast, EGA does not block endosomal recycling of transferrin, retrograde trafficking of ricin, phagolysosomal trafficking, or phagosome permeabilization by Franciscella tularensis. Furthermore, EGA does not neutralize acidic organelles, demonstrating that its mechanism of action is distinct from pH-raising agents such as ammonium chloride and bafilomycin A1. EGA is a powerful tool for the study of membrane trafficking and represents a class of host-targeted compounds for therapeutic development to treat infectious disease.
病原微生物和毒素进化出了多种机制,以进入宿主细胞胞质溶胶,从而对宿主产生毒性作用。一种常见的细胞进入机制需要转运到酸化的内体,这促进了跨宿主膜的易位。为了鉴定阻断该过程的小分子抑制剂,我们筛选了 30,000 种小分子文库,以寻找炭疽致死毒素的抑制剂。在这里,我们报告说,在筛选中发现的最活跃的化合物 4-溴苯甲醛 N-(2,6-二甲基苯基)半卡巴腙,可抑制致死毒素的中毒,并阻止多种其他依赖酸性的细菌毒素和病毒进入哺乳动物细胞。这种化合物,我们称之为 EGA,还延迟了 EGF 受体的溶酶体靶向和降解,表明它靶向宿主膜转运。相比之下,EGA 不会阻断转铁蛋白的内体再循环、蓖麻毒素的逆行运输、吞噬溶酶体运输或土拉弗朗西斯菌的吞噬体穿孔。此外,EGA 不会中和酸性细胞器,表明其作用机制与氯化铵和巴弗洛霉素 A1 等升高 pH 值的试剂不同。EGA 是研究膜转运的有力工具,代表了一类针对宿主的化合物,可用于开发治疗传染病的疗法。