A Selective Irreversible Inhibitor of Furin Does Not Prevent Pseudomonas Aeruginosa Exotoxin A-Induced Airway Epithelial Cytotoxicity.
A Selective Irreversible Inhibitor of Furin Does Not Prevent Pseudomonas Aeruginosa Exotoxin A-Induced Airway Epithelial Cytotoxicity.
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DOI：
10.1371/journal.pone.0159868
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Martin SL
中科院分区：
文献类型：
--
作者：
Ferguson TE;Reihill JA;Walker B;Hamilton RA;Martin SL
许多细菌和病毒病原体(或其毒素)，包括铜绿假单胞菌外毒素A，需要宿主前蛋白转换酶(如呋喃)处理才能致病。我们报道了一种新型的呋喃不可逆抑制剂(QUB-F1)的开发，它包括一个二苯基膦酸亲电弹头和一个底物样肽(RVKR)，RVKR还包括一个生物素标签，以促进基于活性的分析/可视化。与广泛使用的样本化合物(Furin I)相比，QUB-F1对Furin显示出更高的选择性，Furin I具有与RVKR偶联的氯甲基酮弹头，当与丝氨酸胰蛋白酶样酶(胰酶、前列腺素和松解酶)、因子Xa和半胱氨酸蛋白酶组织蛋白酶B进行测试时，QUB-F1对Furin的选择性更高。我们证明QUB-F1不能防止铜绿假单胞菌外毒素A诱导的呼吸道上皮细胞毒性；与Furin I相反，尽管对细胞表面的Furin样活性有类似程度的抑制。这一发现表明，对更广谱的Furin I化合物敏感的其他蛋白酶可能参与了这一过程。
Many bacterial and viral pathogens (or their toxins), including Pseudomonas aeruginosa exotoxin A, require processing by host pro-protein convertases such as furin to cause disease. We report the development of a novel irreversible inhibitor of furin (QUB-F1) consisting of a diphenyl phosphonate electrophilic warhead coupled with a substrate-like peptide (RVKR), that also includes a biotin tag, to facilitate activity-based profiling/visualisation. QUB-F1 displays greater selectivity for furin, in comparison to a widely used exemplar compound (furin I) which has a chloromethylketone warhead coupled to RVKR, when tested against the serine trypsin-like proteases (trypsin, prostasin and matriptase), factor Xa and the cysteine protease cathepsin B. We demonstrate QUB-F1 does not prevent P. aeruginosa exotoxin A-induced airway epithelial cell toxicity; in contrast to furin I, despite inhibiting cell surface furin-like activity to a similar degree. This finding indicates additional proteases, which are sensitive to the more broad-spectrum furin I compound, may be involved in this process.