Penicillin-Binding Protein 5/6 Acting as a Decoy Target in Pseudomonas aeruginosa Identified by Whole-Cell Receptor Binding and Quantitative Systems Pharmacology.
Penicillin-Binding Protein 5/6 Acting as a Decoy Target in Pseudomonas aeruginosa Identified by Whole-Cell Receptor Binding and Quantitative Systems Pharmacology.
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DOI：
10.1128/aac.01603-22
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发表时间：
2023-06-15
影响因子：
4.9
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中科院分区：
文献类型：
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β-内酰胺抗生素已成功用于对抗敏感的铜绿假单胞菌数十年，铜绿假单胞菌具有众所周知的难以穿透外膜（OM）。然而，缺乏关于青霉素结合蛋白（PBP）在完整细菌中对β-内酰胺类和β-内酰胺酶抑制剂的靶位点渗透和共价结合的数据。我们的目的是确定完整和裂解细胞中PBP结合的时间过程，并估计铜绿假单胞菌PAO 1中15种化合物的靶位点渗透和PBP进入。所有β-内酰胺（2 × MIC）均与裂解细菌中的PBPs 1至4显著结合。然而，完整细菌中的PBP结合对于缓慢渗透的β-内酰胺而不是快速渗透的β-内酰胺显著减弱。亚胺培南在1h时产生1.5 ± 0.11 log 10的杀伤作用，而所有其他药物的杀伤作用均<0.5 log 10。相对于亚胺培南，多利培南和美罗培南的净流入率和PBP进入速率慢约2倍，阿维巴坦慢7.6倍，头孢他啶慢14倍，头孢吡肟慢45倍，舒巴坦慢50倍，厄他培南慢72倍，哌拉西林和氨曲南慢约249倍，他唑巴坦慢约358倍，羧苄西林和替卡西林慢约547倍，019倍。在2 × MIC时，PBP 5/6结合程度与净流入率和PBP进入率高度相关（r2 = 0.96），表明PBP 5/6作为诱饵靶标，应通过缓慢渗透、未来的β-内酰胺类药物来避免。这是第一次全面评估完整和裂解的铜绿假单胞菌中PBP结合的时间过程，解释了为什么只有亚胺培南快速杀死。在完整细菌中开发的新型共价结合试验解释了所有表达的耐药机制。
The β-lactam antibiotics have been successfully used for decades to combat susceptible Pseudomonas aeruginosa, which has a notoriously difficult to penetrate outer membrane (OM). However, there is a dearth of data on target site penetration and covalent binding of penicillin-binding proteins (PBP) for β-lactams and β-lactamase inhibitors in intact bacteria. We aimed to determine the time course of PBP binding in intact and lysed cells and estimate the target site penetration and PBP access for 15 compounds in P. aeruginosa PAO1. All β-lactams (at 2 × MIC) considerably bound PBPs 1 to 4 in lysed bacteria. However, PBP binding in intact bacteria was substantially attenuated for slow but not for rapid penetrating β-lactams. Imipenem yielded 1.5 ± 0.11 log10 killing at 1h compared to <0.5 log10 killing for all other drugs. Relative to imipenem, the rate of net influx and PBP access was ~ 2-fold slower for doripenem and meropenem, 7.6-fold for avibactam, 14-fold for ceftazidime, 45-fold for cefepime, 50-fold for sulbactam, 72-fold for ertapenem, ~ 249-fold for piperacillin and aztreonam, 358-fold for tazobactam, ~547-fold for carbenicillin and ticarcillin, and 1,019-fold for cefoxitin. At 2 × MIC, the extent of PBP5/6 binding was highly correlated (r2 = 0.96) with the rate of net influx and PBP access, suggesting that PBP5/6 acted as a decoy target that should be avoided by slowly penetrating, future β-lactams. This first comprehensive assessment of the time course of PBP binding in intact and lysed P. aeruginosa explained why only imipenem killed rapidly. The developed novel covalent binding assay in intact bacteria accounts for all expressed resistance mechanisms.
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影响因子：
6.7
作者：
Lang, Yinzhi;Shah, Nirav R.;Tao, Xun;Reeve, Stephanie M.;Zhou, Jieqiang;Moya, Bartolome;Sayed, Alaa R. M.;Dharuman, Suresh;Oyer, Jeremiah L.;Copik, Alicja J.;Fleischer, Brett A.;Shin, Eunjeong;Werkman, Carolin;Basso, Kari B.;Deveson Lucas, Deanna;Sutaria, Dhruvitkumar S.;Megroz, Marianne;Kim, Tae Hwan;Loudon-Hossler, Victoria;Wright, Amy;Jimenez-Nieves, Rossie H.;Wallace, Miranda J.;Cadet, Keisha C.;Jiao, Yuanyuan;Boyce, John D.;LoVullo, Eric D.;Schweizer, Herbert P.;Bonomo, Robert A.;Bharatham, Nagakumar;Tsuji, Brian T.;Landersdorfer, Cornelia B.;Norris, Michael H.;Soo Shin, Beom;Louie, Arnold;Balasubramanian, Venkataraman;Lee, Richard E.;Drusano, George L.;Bulitta, Juergen B.
通讯作者：
Bulitta, Juergen B.
影响因子：
4.1
作者：
Bergen, Phillip J.;Bulman, Zackery P.;Saju, Sarith;Bulitta, Juergen B.;Landersdorfer, Cornelia;Forrest, Alan;Li, Jian;Nation, Roger L.;Tsuji, Brian T.
通讯作者：
Tsuji, Brian T.
影响因子：
3.2
作者：
Klockgether, Jens;Munder, Antje;Tuemmler, Burkhard
通讯作者：
Tuemmler, Burkhard
影响因子：
3.2
作者：
Dougherty, TJ;Kennedy, K;Pucci, MJ
通讯作者：
Pucci, MJ
影响因子：
4.9
作者：
Li, Henry;Estabrook, Mark;Busha, Karen
通讯作者：
Busha, Karen