Discovery of first-in-class nanomolar inhibitors of heptosyltransferase I reveals a new aminoglycoside target and potential alternative mechanism of action.
Discovery of first-in-class nanomolar inhibitors of heptosyltransferase I reveals a new aminoglycoside target and potential alternative mechanism of action.
复制标题
首次发现纳摩尔级的庚糖基转移酶I抑制剂揭示了一个新的氨基糖苷类靶标和潜在的替代作用机制。
DOI：
10.1038/s41598-022-10776-x
复制
发表时间：
2022-05-04
影响因子：
4.6
通讯作者：
Taylor, Erika A.
中科院分区：
文献类型：
--
作者：
Milicaj, Jozafina;Hassan, Bakar A.;Cote, Joy M.;Ramirez-Mondragon, Carlos A.;Jaunbocus, Nadiya;Rafalowski, Angelika;Patel, Kaelan R.;Castro, Colleen D.;Muthyala, Ramaiah;Sham, Yuk Y.;Taylor, Erika A.
临床相关的Heptosyltransferase I （HepI）抑制剂多年来一直在寻找，因为它在细菌细胞表面脂多糖的生物合成中起着关键作用。虽然许多实验室已经发现或设计了新的小分子抑制剂，但这些化合物缺乏治疗用途所需的生物利用度和效力。HepI蛋白的广泛表征提供了有价值的洞察催化所需的动态运动，可以靶向抑制。kdo2 -脂质A的结构检查提示氨基糖苷类抗生素可能是HepI的潜在抑制剂。多种氨基糖苷已被实验证实为HepI的纳米摩尔抑制剂，其中最佳抑制剂Ki为600±90 nM。通过详细的动力学分析来确定抑制机制，同时利用圆二色光谱、固有色氨酸荧光、对接和分子动力学模拟来证实动力学实验结果。虽然氨基糖苷长期以来被描述为针对细菌核糖体蛋白质合成的强效抗生素，导致细菌细胞膜稳定性的破坏，但最近研究人员表明，它们对蛋白质生产的影响很小。我们的研究提示了氨基糖苷类化合物抑制HepI的另一种新的作用机制，它直接导致体内LPS产生的改变。这一发现可能会改变我们对氨基糖苷类抗生素功能的理解，因为脂多糖生物合成的中断是氨基糖苷作用的另一个重要机制。进一步研究氨基糖苷对细胞的微生物影响是有必要的，因为抑制核糖体可能不是唯一和主要的作用机制。氨基糖苷对HepI的抑制作用可能会显著改变修饰氨基糖苷结构的策略，从而提高抗细菌感染的效果。
A clinically relevant inhibitor for Heptosyltransferase I (HepI) has been sought after for many years because of its critical role in the biosynthesis of lipopolysaccharides on bacterial cell surfaces. While many labs have discovered or designed novel small molecule inhibitors, these compounds lacked the bioavailability and potency necessary for therapeutic use. Extensive characterization of the HepI protein has provided valuable insight into the dynamic motions necessary for catalysis that could be targeted for inhibition. Structural inspection of Kdo2-lipid A suggested aminoglycoside antibiotics as potential inhibitors for HepI. Multiple aminoglycosides have been experimentally validated to be first-in-class nanomolar inhibitors of HepI, with the best inhibitor demonstrating a Ki of 600 ± 90 nM. Detailed kinetic analyses were performed to determine the mechanism of inhibition while circular dichroism spectroscopy, intrinsic tryptophan fluorescence, docking, and molecular dynamics simulations were used to corroborate kinetic experimental findings. While aminoglycosides have long been described as potent antibiotics targeting bacterial ribosomes’ protein synthesis leading to disruption of the stability of bacterial cell membranes, more recently researchers have shown that they only modestly impact protein production. Our research suggests an alternative and novel mechanism of action of aminoglycosides in the inhibition of HepI, which directly leads to modification of LPS production in vivo. This finding could change our understanding of how aminoglycoside antibiotics function, with interruption of LPS biosynthesis being an additional and important mechanism of aminoglycoside action. Further research to discern the microbiological impact of aminoglycosides on cells is warranted, as inhibition of the ribosome may not be the sole and primary mechanism of action. The inhibition of HepI by aminoglycosides may dramatically alter strategies to modify the structure of aminoglycosides to improve the efficacy in fighting bacterial infections.
登录
查看更多内容
DOI：
10.1007/978-1-62703-245-2_15
发表时间：
2013
期刊：
Methods in molecular biology (Clifton, N.J.)
影响因子：
--
作者：
Hankins JV;Madsen JA;Needham BD;Brodbelt JS;Trent MS
通讯作者：
Trent MS
影响因子：
2.9
作者：
Czyzyk, Daniel J.;Liu, Cassie;Taylor, Erika A.
通讯作者：
Taylor, Erika A.
DOI：
10.1073/pnas.83.16.6164
发表时间：
1986-08-01
影响因子：
11.1
作者：
DAVIS, BD;CHEN, LL;TAI, PC
通讯作者：
TAI, PC
影响因子：
2.9
作者：
Cote, Joy M.;Ramirez-Mondragon, Carlos A.;Taylor, Erika A.
通讯作者：
Taylor, Erika A.
影响因子：
3.2
作者：
HITCHCOCK, PJ;BROWN, TM
通讯作者：
BROWN, TM