STRUCTURE BIOLOGY OF ORAL BACTERIAL PROTEINS THAT ERADICATE BIOFILMS

消除生物膜的口腔细菌蛋白的结构生物学

• 批准号: 8171531
• 负责人: NARAYANAN RAMASUBBU
• 金额: $ 1.43万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-01 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8171531
• 关键词: Acetylglucosamine; Architecture; Bacterial Proteins; Catalytic Domain; Cleaved cell; Complex; Computer Retrieval of Information on Scientific Projects Database; Data; Data Collection; Deacetylase; Distal; Enzymes; Excision; Funding; Grant; Housing; Institution; Ligands; Link; Maps; Microbial Biofilms; Mutation; Oral; Polymers; Research; Research Personnel; Resolution; Resources; Site; Source; Staphylococcus aureus; Structure; United States National Institutes of Health; Work; inhibitor/antagonist; mutant; oral biology

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. This will be a continuation of previous projects that the PI has submitted and gotten beamtime. Briefly, we are working on enzymes that degradde and detach oral as well as meedically important biofilms. The bacterial species that produce these biofilms include Aggregatibacter actinomycetemcomitans, S. epidermidis and S. aureus. We are studying two enzymes, Dispersin B and PgaB. Both work on the ssubstrate, a homopolymer of beta(1,6)linked N-acetylglucosamine. While dispersin B cleaves the beta(1,6 linkages, PgaB deacetylates the N-acetylglucosamine groups of the polymer. Both sensitize each other in efficient removal of biofilms. We have obtained crystals of Dispersin B that diffract to 2.0 A in house. We would like to collect high resolution data for crystals that are soaked with substrate and inhibitors. The complex also diffracts to 2 A, but we are unable to resolve the ligand well in the maps. We would like to use the high resolution data to see whether this can be done. Four to 5 mutants will be brought for data collection. Some of the mutations occur at the catalytic site and others at distal sites. The hypothesis is that altough dispersin B is an exo enzyme, it does have multiple subsites. Resolution of subsite architecture will also be analyzed. The second enzyme is a deacetylase and we have good crystals that diffract to 8 A. We would like to see whether good, usuable data can be obtained.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目和 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 这将是 PI 已提交并获得 Beamtime 的先前项目的延续。简而言之，我们正在研究能够降解和分离口腔以及医学上重要的生物膜的酶。产生这些生物膜的细菌种类包括放线菌聚集杆菌、表皮葡萄球菌和金黄色葡萄球菌。我们正在研究两种酶：Dispersin B 和 PgaB。两者都作用于 s 底物，即 β(1,6) 连接的 N-乙酰氨基葡萄糖的均聚物。分散素 B 裂解 β(1,6 连接)，而 PgaB 使聚合物的 N-乙酰葡糖胺基团脱乙酰基。两者在有效去除生物膜时相互敏化。 我们已经获得了内部衍射度为 2.0 A 的分散素 B 晶体。我们希望收集用底物和抑制剂浸泡的晶体的高分辨率数据。该配合物也衍射至 2 A，但我们无法在图中很好地解析配体。我们想使用高分辨率数据来看看这是否可以做到。将带四到五个突变体进行数据收集。一些突变发生在催化位点，另一些突变发生在远端位点。假设虽然分散素 B 是一种外切酶，但它确实具有多个亚位点。子站点架构的分辨率也将被分析。 第二种酶是脱乙酰酶，我们有衍射至 8 A 的良好晶体。我们想看看是否可以获得好的、可用的数据。