CRYSTALLOGRAPHIC STUDIES OF ANTIBIOTIC RESISTANCE PROTEINS AND SIGNAL TRANSDUCTI

抗生素耐药蛋白和信号转导的晶体学研究

• 批准号: 8362188
• 负责人: FOCCO VAN DEN AKKER
• 金额: $ 0.14万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-03-01 至 2012-02-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8362188
• 关键词: Antibiotic Resistance; Antibiotics; Bacterial Infections; Binding; Bone Growth; Carbapenems; Clinical Trials; Coiled-Coil Domain; Collaborations; Complex; Cyclic Nucleotides; Data Set; Development; Disease Outbreaks; Funding; Grant; Guanylate Cyclase; Human; Ion Channel; Klebsiella pneumonia bacterium; L-Selenomethionine; Lactamase; Lead; Link; Methodist Church; Molecular; National Center for Research Resources; New York; Pharmacologic Substance; Physiological Processes; Principal Investigator; Radiation; Research; Research Infrastructure; Resolution; Resort; Resources; Roentgen Rays; Signal Pathway; Signal Transduction; Signaling Protein; Source; Structure; United States National Institutes of Health; Universities; beta-lactamase PSE-2; blood pressure regulation; clinically relevant; cost; design; inhibitor/antagonist; insight; novel; structural biology; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. The development of antibiotic resistance to bacterial infections is a serious human threat in large part due to bacterial b-lactamases. Inhibition of these b-lactamases is therefore a key pharmaceutical approach. Our lab focuses on delineating the molecular inhibition mechanism of clinically available inhibitors, additional potent inhibitors that are in, or close to, clinical trials, and our own designed inhibitors. The inhibition mechanism is complex involving a large number of covalent intermediates that we study using a novel X-ray and Raman crystallographic approach. High resolution crystallographic studies are proposed for inhibitor complexes for a variety of different clinically relevant b-lactamases classes including KPC-2, recently linked to an K. pneumoniae outbreak in New York, SHV-1, and OXA-1, OXA-10, and OXA-24/40. Both OXA24/40 and KPC-2 are a major threat to carbapenems, a last resort antibiotic. Our inhibitors are developed in collaboration with Dr. Buynak (Southern Methodist University) and are designed to form a stable inhibitory intermediate, either by forming a trans-enamine or bicyclic aromatic ring intermediate. In addition, we are embarking on using fragment-mixture soaked b-lactamase crystals as a tool for finding new lead compounds. In addition, our lab focuses on structural studies of cyclic nucleotide signaling. We have recently crystallized the coiled-coil domain of a guanylyl cyclase involved in blood pressure regulation. Crystals diffract to 2¿ and we plan on collecting a SeMet MAD dataset. Finally, we have crystallized several cyclic nucleotide binding domains of ion channels of which we determined the structure of one, and have diffracting crystals for two others. The latter project is aimed to providing structural insights into cyclic nucleotide signaling pathways involving in blood pressure regulation, bone growth and other important physiological processes.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供 该子项目的主要支持。 并且子项目的首席研究员可能是由其他来源提供的， 包括其他 NIH 来源的子项目可能列出的总成本。 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 细菌感染的抗生素耐药性的发展对人类构成了严重威胁，这在很大程度上是由于细菌 β-内酰胺酶造成的，因此，我们的实验室重点研究临床可用抑制剂的分子抑制机制。正在进行或接近临床试验的其他有效抑制剂，以及我们自己设计的抑制剂，其抑制机制很复杂，涉及大量共价中间体，我们使用新型 X 射线和拉曼高分辨率晶体学方法进行研究。建议对各种不同临床相关 β-内酰胺酶类别的抑制剂复合物进行晶体学研究，包括最近与纽约肺炎克雷伯菌爆发有关的 KPC-2、SHV-1 和 OXA-1、OXA-10 和 OXA -24/40。OXA24/40 和 KPC-2 都是碳青霉烯类抗生素的主要威胁，我们的抑制剂是与 Buynak 博士合作开发的。 （南卫理公会大学），旨在通过形成反式烯胺或双环芳环中间体来形成稳定的抑制中间体。此外，我们正着手使用片段混合物浸泡的 β-内酰胺酶晶体作为寻找新的工具。此外，我们的实验室专注于环核苷酸信号传导的结构研究，最近我们结晶了参与血压调节的鸟苷酸环化酶晶体。 2¿我们计划收集 SeMet MAD 数据集，最后，我们确定了离子通道的几个环核苷酸结合域的结构，并获得了另外两个的衍射晶体。后一个项目旨在提供结构见解。环核苷酸信号通路涉及血压调节、骨骼生长和其他重要的生理过程。