Inhibition of HeptosyltransferaseI for the treatment of Gram-negative bacterial infection

抑制庚基转移酶 I 治疗革兰氏阴性菌感染

基本信息

批准号：
8958425
负责人：
Erika A Taylor
金额：
$ 49.29万
依托单位：
WESLEYAN UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-06-15 至 2019-06-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8958425
关键词：
Address Affinity Anabolism Area Bacteria Binding Biochemical Biochemistry Biological Process Biophysics Carbon Catalysis Cell surface Cessation of life Charge Chemicals Chemistry Computer Analysis Data Deuterium Development Docking Drug Design Ensure Environment Enzymes Escherichia coli Fluorescence Fluorescent Probes Geometry Goals Gram-Negative Bacteria Gram-Negative Bacterial Infections Hydrogen Intestines Investigation Isotopes Kinetics Laboratories Lead Libraries Ligand Binding Lipopolysaccharide Biosynthesis Pathway Lipopolysaccharides Mechanics Membrane Microbial Biofilms Microbial Drug Resistance Molecular Bank Molecular Structure Motion Nature Process Protein Dynamics Proteins Reaction Reporting Research Running Sampling Science Structural Protein Structure Students Swimming System Systems Development Techniques Therapeutic Therapeutic Agents Time Training Underrepresented Students Universities Work analog antimicrobial base cell motility design drug development drug discovery glycosylation glycosyltransferase graduate student improved inhibitor/antagonist protein B protein structure function public health relevance quantum small molecule stereochemistry stopped-flow fluorescence success sugar undergraduate student

项目摘要

DESCRIPTION (provided by applicant): With numerous Gram-negative bacterial species demonstrating antimicrobial drug resistance, the identification of new targets for inhibitor design in bacterial systems is of great importance. Escherichia coli, common Gram-negative bacteria, cause illness in a quarter of a million people and hundreds of deaths each year, in the US alone. As part of our goal of helping to develop new antimicrobial compounds, we have been investigating the heptosyltransferases involved in the biosynthesis of the core region of the lipopolysaccharide (LPS) from Escherichia coli. Heptosyltransferase (Hep) enzymes are essential for the formation of bacterial biofilms in Gram-negative bacteria, making the Hep enzymes an important targets for the development of biofilm inhibitors. Our lab recently demonstrated that E. coli HepI is the first LPS biosynthetic enzyme capable of utilizing fully delipidated substrate analogues while maintaining enzymatic proficiency. Additionally, our work has revealed time-resolved protein dynamics for HepI. This data, along with HepI structural information make this a promising system for the development of inhibitors. Our investigation will address two hypotheses: (1) inhibitor design for this and other glycosyltransferases (GTs) can be improved through characterization of the HepI transition state by kinetic isotope effect studies, and (2) that HepI undergoes significant conformational changes upon ligand binding, the disruption of which might be useful for development of inhibitors for HepI and also other GTs. To date, many inhibitors have been developed for glycosyltransferase enzymes; however, these inhibitors are typically not sufficiently tight-binding for drug development. This proposal seeks to determine whether HepI catalyzes a SN1- or SN2-like reaction in order to enhance our ability to design potent inhibitors for this important class of enzymes. Additionally, since in som enzymological systems including HepI, protein dynamics are necessary for chemistry, efforts to identify inhibitors that not only compete with the substrates, but also those that can disrupt protein dynamics are being pursued. We have recently reported the first transient kinetic analyses on a GT-B protein using stopped-flow fluorescence analyses, and we plan to continue these investigations to allow for determination of whether our inhibitors disrupt chemistry or protein structural changes, like those observed in crystal structures of GTs of the GT-B class. This work promises to enhance drug discovery efforts for multiple systems, including the inhibition of bacterial biofilm formation through inhibition of HepI. 1

描述（由应用提供）：有大量革兰氏阴性细菌具有抗菌耐药性，鉴定了抑制剂设计的新靶在细菌中，系统非常重要。大肠杆菌（Escherichia Coli）是普通的革兰氏阴性细菌，仅在美国，每年在四分之一的人中会导致四分之一的人死亡。作为帮助开发新的抗微生物化合物的目标的一部分，我们一直在研究与大肠杆菌脂多糖（LPS）核心区域的生物合成有关的七糖基转移酶。七糖基转移酶（HEP）酶对于革兰氏阴性细菌中细菌生物膜的形成至关重要，这使得HEP酶是生物抑制剂发展的重要靶标。我们的实验室最近证明，大肠杆菌HEPI是第一个能够利用完全部署的底物类似物的LPS生物合成酶，同时保持酶促的熟练度。此外，我们的工作揭示了HEPI的时间分辨蛋白质动力学。 This data, along with HepI structural information make this a promising system for the Our investment will address two hypotheses: (1) inhibitor design for this and other glycosyltransferases (GTs) can be improved through characterization of the HepI transition state by kinetic isotope effect studies, and (2) that HepI undergoes significant conformational changes upon ligand binding, the disruption of which might be useful for development of inhibitors对于HEPI以及其他GTS。迄今为止，已经开发了许多用于糖基转移酶的抑制剂。但是，这些抑制剂通常没有足够的紧密结合来开发药物。该提案旨在确定HEPI是否会催化SN1或SN2样反应，以增强我们为这类酶设计潜在抑制剂的能力。此外，由于在包括HEPI在内的SOM酶学系统中，蛋白质动力学对于化学是必要的，努力识别不仅与底物竞争的抑制剂，而且还需要破坏蛋白质动力学的抑制剂。我们最近报告了使用停止流量荧光分析对GT-B蛋白进行的首次短暂动力学分析，我们计划继续这些投资，以确定我们的抑制剂是否破坏化学或蛋白质结构变化，例如GT-B类GTS晶体结构中观察到的抑制剂。这项工作有望增强多种系统的药物发现工作，包括通过抑制HEPI抑制生物膜形成。 1

项目成果

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Ligand-Induced Conformational and Dynamical Changes in a GT-B Glycosyltransferase: Molecular Dynamics Simulations of Heptosyltransferase I Complexes.

DOI：
10.1021/acs.jcim.1c00868
发表时间：
2022-01-24
期刊：
Journal of chemical information and modeling
影响因子：
5.6
作者：
Hassan BA;Milicaj J;Ramirez-Mondragon CA;Sham YY;Taylor EA
通讯作者：
Taylor EA

The Glycosyltransferases of LPS Core: A Review of Four Heptosyltransferase Enzymes in Context.

LPS 核心的糖基转移酶：四种肽基转移酶的背景综述。

DOI：
10.3390/ijms18112256
发表时间：
2017-10-27
期刊：
International journal of molecular sciences
影响因子：
5.6
作者：
Cote JM;Taylor EA
通讯作者：
Taylor EA

Conserved Conformational Hierarchy across Functionally Divergent Glycosyltransferases of the GT-B Structural Superfamily as Determined from Microsecond Molecular Dynamics.

DOI：
10.3390/ijms22094619
发表时间：
2021-04-28
期刊：
International journal of molecular sciences
影响因子：
5.6
作者：
Ramirez-Mondragon CA;Nguyen ME;Milicaj J;Hassan BA;Tucci FJ;Muthyala R;Gao J;Taylor EA;Sham YY
通讯作者：
Sham YY

Synthesis, kinetics and inhibition of Escherichia coli Heptosyltransferase I by monosaccharide analogues of Lipid A.

脂质 A 的单糖类似物对大肠杆菌庚糖基转移酶 I 的合成、动力学和抑制作用。

DOI：
10.1016/j.bmcl.2018.01.040
发表时间：
2018
期刊：
Bioorganic & medicinal chemistry letters
影响因子：
2.7
作者：
Nkosana,NoreenK;Czyzyk,DanielJ;Siegel,ZarekS;Cote,JoyM;Taylor,ErikaA
通讯作者：
Taylor,ErikaA

Discovery of first-in-class nanomolar inhibitors of heptosyltransferase I reveals a new aminoglycoside target and potential alternative mechanism of action.

DOI：
10.1038/s41598-022-10776-x
发表时间：
2022-05-04
期刊：
SCIENTIFIC REPORTS
影响因子：
4.6
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.
通讯作者：
Taylor, Erika A.