STRUCTURAL STUDIES ON AMINOGLYCOSIDE PHOSPHOTRANSFERASES

氨基糖苷磷酸转移酶的结构研究

• 批准号: 8169967
• 负责人: CLYDE A SMITH
• 金额: $ 0.27万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-05-01 至 2011-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169967
• 关键词: Aminoglycoside resistance; Aminoglycosides; Antibiotic Resistance; Antibiotics; Binding; Complex; Computer Retrieval of Information on Scientific Projects Database; Crystallization; Data; Enterococcus; Enterococcus faecium; Enzyme Inhibitor Drugs; Enzyme Inhibitors; Enzymes; Family; Funding; Gentamicins; Grant; Institution; Kanamycin; Kanamycin Kinase; L-Selenomethionine; Mediating; Modeling; Molecular; Multienzyme Complexes; Phosphotransferases; Publishing; Reporting; Research; Research Personnel; Resistance; Resources; Source; Streptomycin; Structure; Tobramycin; United States National Institutes of Health; design; hydroxyl group; three dimensional structure

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. A major factor in the emergence of antibiotic resistance is the existence of bacterial enzymes that chemically modify common antibiotics. One such family of anti-bacterials to which there is now almost universal resistance are the aminoglycosides (e.g. kanamycin, tobramycin and gentimicin). High level resistance to gentamicin in enterococci is mediated by a group of four phosphotransferases belonging to the APH(2?) sub-family of enzymes which phosphorylate at a specific hydroxyl group on the antibiotic, using ATP as a cosubstrate. An understanding of how these enzymes bind and deactivate the aminoglycosides will provide valuable information for the design of specific inhibitors of these enzymes. We are studying all four of the APH(2?) phosphotransferases, APH(2?)-Ia and APH(2?)-IIa from Enterococcus faecium, APH(2?)-IIIa from E. gallinarum, and APH(2?)-IVa from E. casseliflavus. The three dimensional structures of the binary gentamicin complex and a ternary AMPPCP-streptomycin complex of APH(2?)-IIa has been determined and recently published and the other crystallization conditions of APH(2?)-IIIa and APH(2?)-IVa have been reported. Both the APH(2?)-IIIa and APH(2?)-IVa structures has been solved by molecular replacement using APH(2?)-IIa as the search model. In addition, three wavelength MAD data has been collected from a SeMet APH(2?)-Ia crystal and the structure has been solved and partially refined. Additional structural studies on binary and ternary complexes of these enzymes are underway.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 抗生素耐药性出现的一个主要因素是存在化学修饰常见抗生素的细菌酶。氨基糖苷类抗生素（如卡那霉素、妥布霉素和庆大霉素）就是目前几乎普遍耐药的一类抗菌药物。肠球菌对庆大霉素的高水平耐药是由APH（2？）在抗生素上的特定羟基上磷酸化的酶的亚家族，使用ATP作为共底物。了解这些酶如何结合和失活的氨基糖苷类将提供有价值的信息，这些酶的特异性抑制剂的设计。 我们正在研究所有四个APH（2？）磷酸转移酶，APH（2？）- Ia和APH（2？）- IIa来自屎肠球菌，APH（2？）- IIIa来自E. gallinarum和APH（2？）IVa来自E.石蒜黄APH（2？）-庆大霉素二元复合物和AMPPCP-链霉素三元复合物的三维结构IIa已被确定并最近发表，APH（2？）- IIIa和APH（2？）- IVa已报告。 APH（2？）- IIIa和APH（2？）- IVa结构已经通过使用APH（2？）- IIa作为搜索模型。 此外，还从SeMet APH（2？）Ia晶体，结构已得到解决和部分完善。 这些酶的二元和三元复合物的其他结构研究正在进行中。