Identification of Staphylococcus aureus Penicillin Binding Protein 4 (PBP4) Inhibitors.
Identification of Staphylococcus aureus Penicillin Binding Protein 4 (PBP4) Inhibitors.
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DOI：
10.3390/antibiotics11101351
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发表时间：
2022-10-04
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影响因子：
--
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耐甲氧西林金黄色葡萄球菌（MRSA）是一个全球性的医疗保健问题。这种耐药性历来归因于获得mecA（或mecC），其编码具有低β-内酰胺亲和力的替代青霉素结合蛋白PBP 2a。然而，最近的研究表明，青霉素结合蛋白4（PBP 4）也是S。金黄色葡萄球菌甲氧西林耐药性，特别是社区获得性MRSA菌株。因此，PBP 4被认为是一个有趣的治疗靶点，因为相应的抑制剂可以恢复MRSA β-内酰胺敏感性。除了在抗生素耐药性中的作用外，PBP 4最近也被证明是S.金黄色葡萄球菌皮质骨骨细胞腔隙-小管网络（OLCN）的侵袭和定植，为骨感染的复发提供了一个生态位。从这些角度来看，PBP 4抑制剂的发展可能会产生巨大的影响，作为逆转甲氧西林耐药性和抑制生物体引起慢性骨髓炎的能力的药物。因此，使用30，000个成员的小分子化学文库的全细胞高通量筛选和二级测定，我们鉴定了推定的S。金黄色葡萄球菌PBP 4抑制剂。定量逆转录酶介导的PCR和PBP 4结合测定显示，命中可以进一步区分为降低PBP 4表达的化合物与可能影响蛋白质功能的化合物。我们还发现，6.25 µM（2.5 µg/mL）的主要候选药物9314848可逆转微生物的PBP 4依赖性MRSA表型，并抑制其穿过模拟OLCN孔的微流控硅膜小管阵列（µSiM-CA）的能力。总的来说，这些分子可能代表作为PBP 4抑制剂的有希望的潜力，其可以进一步开发为用于治疗MRSA感染和/或骨髓炎的佐剂。
Methicillin-resistant Staphylococcus aureus (MRSA) is a global healthcare concern. Such resistance has historically been attributed to the acquisition of mecA (or mecC), which encodes an alternative penicillin binding protein, PBP2a, with low β-lactam affinity. However, recent studies have indicated that penicillin binding protein 4 (PBP4) is also a critical determinant of S. aureus methicillin resistance, particularly among community-acquired MRSA strains. Thus, PBP4 has been considered an intriguing therapeutic target as corresponding inhibitors may restore MRSA β-lactam susceptibility. In addition to its role in antibiotic resistance, PBP4 has also recently been shown to be required for S. aureus cortical bone osteocyte lacuno-canalicular network (OLCN) invasion and colonization, providing the organism with a niche for re-occurring bone infection. From these perspectives, the development of PBP4 inhibitors may have tremendous impact as agents that both reverse methicillin resistance and inhibit the organism’s ability to cause chronic osteomyelitis. Accordingly, using a whole-cell high-throughput screen of a 30,000-member small molecule chemical library and secondary assays we identified putative S. aureus PBP4 inhibitors. Quantitative reverse transcriptase mediated PCR and PBP4 binding assays revealed that hits could be further distinguished as compounds that reduce PBP4 expression versus compounds that are likely to affect the protein’s function. We also showed that 6.25 µM (2.5 µg/mL) of the lead candidate, 9314848, reverses the organism’s PBP4-dependent MRSA phenotype and inhibits its ability to traverse Microfluidic-Silicon Membrane-Canalicular Arrays (µSiM-CA) that model the OLCN orifice. Collectively, these molecules may represent promising potential as PBP4-inhibitors that can be further developed as adjuvants for the treatment of MRSA infections and/or osteomyelitis prophylactics.
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影响因子：
4.9
作者：
Chatterjee, Som S.;Chen, Liang;Chambers, Henry F.
通讯作者：
Chambers, Henry F.
影响因子：
56.3
作者：
Garcia-Alvarez, Laura;Holden, Matthew T. G.;Lindsay, Heather;Webb, Cerian R.;Brown, Derek F. J.;Curran, Martin D.;Walpole, Enid;Brooks, Karen;Pickard, Derek J.;Teale, Christopher;Parkhill, Julian;Bentley, Stephen D.;Edwards, Giles F.;Girvan, E. Kirsty;Kearns, Angela M.;Pichon, Bruno;Hill, Robert L. R.;Larsen, Anders Rhod;Skov, Robert L.;Peacock, Sharon J.;Maskell, Duncan J.;Holmes, Mark A.
通讯作者：
Holmes, Mark A.
影响因子：
5.3
作者：
Kremers, Hilal Maradit;Nwojo, Macaulay E.;Huddleston, Paul M., III
通讯作者：
Huddleston, Paul M., III
DOI：
10.1073/pnas.74.3.1009
发表时间：
1977-01-01
影响因子：
11.1
作者：
GEORGOPAPADAKOU, N;HAMMARSTROM, S;STROMINGER, JL
通讯作者：
STROMINGER, JL
影响因子：
3.4
作者：
Loskill, Peter;Pereira, Pedro M.;Jacobs, Karin
通讯作者：
Jacobs, Karin