Recombinant HNP-1 Produced by Escherichia coli Triggers Bacterial Apoptosis and Exhibits Antibacterial Activity against Drug-Resistant Bacteria.
Recombinant HNP-1 Produced by Escherichia coli Triggers Bacterial Apoptosis and Exhibits Antibacterial Activity against Drug-Resistant Bacteria.
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大肠杆菌产生的重组 HNP-1 触发细菌凋亡并对耐药细菌表现出抗菌活性
DOI：
10.1128/spectrum.00860-21
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发表时间：
2022-02-23
影响因子：
3.7
通讯作者：
Xu P
中科院分区：
文献类型：
--
作者：
Xie Q;Wang Y;Zhang M;Wu S;Wei W;Xiao W;Wang Y;Zhao J;Liu N;Jin Y;Wu J;Xu P
人中性粒细胞肽-1 (HNP-1) 是一种很有前途的候选抗生素，但其临床应用因大规模生产过程中的挑战以及对其杀菌机制了解不足而受到阻碍。在本研究中，我们证明表达全长 preproHNP-1 的大肠杆菌分泌可溶形式的 HNP-1，可在异丙硫基-β-d-半乳糖苷 (IPTG) 诱导和超滤后从总细胞裂解液中回收。无标记定量蛋白质组学和免疫共沉淀实验表明，HNP-1 通过引起 DNA 和膜损伤来诱导细菌细胞凋亡。值得注意的是，我们发现 HNP-1 通过干扰 RecA 与单链 DNA (ssDNA) 的结合来破坏 DNA 损伤反应途径。进一步的实验表明，封装在脂质体中的HNP-1可抑制耐甲氧西林金黄色葡萄球菌（MRSA）和耐美罗培南铜绿假单胞菌（MRPA）的生长。这些结果表明重组蛋白表达可能是生产 HNP-1 的简单且经济高效的解决方案，并且通过 HNP-1 抑制 RecA 可以作为对抗抗生素耐药性的替代策略。重要性 人中性粒细胞肽-1 (HNP-1) 是一种有前途的候选抗生素，但其临床应用因大规模生产的困难和对其杀菌机制了解不足而受到阻碍。在这项研究中，我们证明重组蛋白表达与超滤相结合可能是 HNP-1 生产的一种简单且经济高效的解决方案。我们进一步发现HNP-1诱导细菌凋亡并阻止其SOS修复途径与RecA蛋白结合，这可能是一种新的抗菌机制。此外，我们还发现，封装在脂质体中的 HNP-1 可抑制耐甲氧西林金黄色葡萄球菌 (MRSA) 和耐美罗培南铜绿假单胞菌 (MRPA) 的生长。这些结果为HNP-1的产生和抗菌机制提供了新的见解，这两者都可能促进其临床应用。
Human neutrophil peptide-1 (HNP-1) is a promising antibiotic candidate, but its clinical applications have been hampered by challenges during mass production and an inadequate understanding of its bactericidal mechanisms. In this study, we demonstrated that Escherichia coli expressing full-length preproHNP-1 secretes a soluble form of HNP-1, which can be recovered from the total cell lysate after isopropyl thio-β-d-galactoside (IPTG) induction and ultrafiltration. Label-free quantitative proteomics and co-immunoprecipitation experiments revealed that HNP-1 induces cell apoptosis in bacteria by causing DNA and membrane damage. Notably, we found that HNP-1 disrupts the DNA damage response pathway by interfering with the binding of RecA to single-stranded DNA (ssDNA). Further experiments demonstrated that HNP-1 encapsulated in liposomes inhibits the growth of methicillin-resistant Staphylococcus aureus (MRSA) and meropenem-resistant Pseudomonas aeruginosa (MRPA). These results indicated that recombinant protein expression may be a simple and cost-effective solution to produce HNP-1 and that RecA inhibition via HNP-1 may serve as an alternative strategy to counteract antibiotic resistance. IMPORTANCE Human neutrophil peptide-1 (HNP-1) is a promising antibiotic candidate, but its clinical application has been hampered by the difficulty of mass production and an inadequate understanding of its bactericidal mechanisms. In this study, we demonstrated that recombinant protein expression combined with ultrafiltration may be a simple and cost-effective solution to HNP-1 production. We further found that HNP-1 induces bacterial apoptosis and prevents its SOS repair pathway from binding to the RecA protein, which may be a new antibacterial mechanism. In addition, we showed that HNP-1 encapsulated in liposomes inhibits the growth of methicillin-resistant Staphylococcus aureus (MRSA) and meropenem-resistant Pseudomonas aeruginosa (MRPA). These results provide new insights into the production and antibacterial mechanism of HNP-1, both of which may promote its clinical application.
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影响因子：
1.8
作者：
Larentis AL;Nicolau JF;Esteves Gdos S;Vareschini DT;de Almeida FV;dos Reis MG;Galler R;Medeiros MA
通讯作者：
Medeiros MA
影响因子：
64.5
作者：
Kohanski, Michael A.;Dwyer, Daniel J.;Collins, James J.
通讯作者：
Collins, James J.
影响因子：
1.6
作者：
Pazgier, Marzena;Lubkowski, Jacek
通讯作者：
Lubkowski, Jacek
影响因子：
5.8
作者：
Li, Miao;Zhang, Tongtong;Jin, Yiguang
通讯作者：
Jin, Yiguang
影响因子：
5.2
作者：
El Shazely, Baydaa;Yu, Guozhi;Rolff, Jens
通讯作者：
Rolff, Jens