HIGH RESOLUTION DATA SET OF 2 FORMS OF VANCOMYCIN RESISTANCE ACCOCIATED

两种形式的万古霉素耐药性的高分辨率数据集

• 批准号: 7721327
• 负责人: EMIL PAI
• 金额: $ 2.09万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721327
• 关键词: Active Sites; Anabolism; Anodes; Archaea; Bacteria; Binding; Cell Wall; Cleaved cell; Computer Retrieval of Information on Scientific Projects Database; Consensus; Data; Data Set; Enzymes; Escherichia coli; Freezing; Funding; Grant; Institution; Mammals; Peptides; Peptidoglycan; Pharmaceutical Preparations; Proteins; Research; Research Personnel; Resolution; Resources; Serine Protease; Source; Standards of Weights and Measures; Structure; Temperature; Testing; Time; United States National Institutes of Health; Vancomycin; Vancomycin Resistance; Work; inorganic phosphate; mutant; response

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. 1) Vancomycin resistance accociated Regulator (VraR) is involved in mounting a response to antibiotices that target cell wall peptidoglycan biosynthesis such as vancomycin. This enzyme is not present in mammals and thus becomes a good target for a drug to target vancomycin resistant bacteria. (2 MAD data sets + 2 high resolution) (1 MAD set and high res is absolutely needed now, the highest priority) The other MAD set and high res is the second thing that we need from this trip. Also, 2 high resolution sets of a different space group would be collected last on the trip. Only if there is extra time, this is not a primary data set. All of these crystals have been frozen and pre-screened on our rotating anode --------------------------------------------------------------- 2) E. coli serine protease (ECSP) cleaves several significant proteins. We have previously solved the structure and would now like to look at consensus peptides bound to the active site. 2 data sets - The 2.5A was low exposure just to check the crystal quality of pre-frzen crystals. Crystals are ordered to better than 2A. All of these crystals have been frozen and pre-screened on our rotating anode. These will diffract to higher resolution at CHESS. The native protein diffracts much better, better than 2A. We are trying to get some additional similar standard peptides into the protein. If these work before the trip, we will pre-freeze and pre-screen them and bring them along to collect. -------------------------------------------------------------- 3) (S)-3-O-Geranylgeranlyglyceryl Phosphate Synthase (GGGPS) is isolated from Archaea and is involved in synthesis of compounds unique to Archaea cell walls. We would like to look at the mechanism of this unique type of enzymes. 3 data sets - they have been frozen and pre-screened on our rotating anode and stored for the trip. There are a few other similar mutants being crystallized. If we can get crystals, pre-freeze and pre-test them, we will bring them along to collect, if there is time. ------------------------------------------------------------------ Every crystal on our list has been pre-frozen and pre-screened on our rotating anode. We will bring some room temperature plates, but purely as a backup. The MAD data on VraR is our priority for this trip.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 1)万古霉素抗性相关调节因子(VRAR)参与了对针对细胞壁肽聚糖生物合成的抗生素(如万古霉素)的反应。这种酶在哺乳动物中不存在，因此成为针对万古霉素耐药细菌的药物的良好靶点。(2个MAD数据集+2个高分辨率)(现在绝对需要1个MAD集和高分辨率，最高优先级)另一个MAD集和高分辨率是我们此行需要的第二件事。另外，两组不同空间组的高分辨率图像将在旅程的最后一次收集。只有在有额外时间的情况下，这才不是主要数据集。所有这些晶体都被冷冻，并在我们的旋转阳极上进行预筛选。---------------------------------------------------------------2)大肠杆菌丝氨酸蛋白酶能裂解几种重要的蛋白质。我们之前已经解决了结构问题，现在想看看与活性部位结合的共识多肽。2个数据集-2.5A是低曝光量，只是为了检查冷冻前晶体的晶体质量。晶体被排序到比2a更好。所有这些晶体都被冻结，并在我们的旋转阳极上进行预筛选。这些将在国际象棋中转化为更高的分辨率。天然蛋白质的衍射率要好得多，比2A好。我们正试图在蛋白质中加入更多类似的标准多肽。如果这些在旅行前有效，我们将对它们进行预冻和预筛选，并将它们随身带着收集。--------------------------------------------------------------3)(S)-3-O-香叶基香叶甘油基磷酸合成酶(GGGPS)是从古生代植物中分离得到的，参与古生代植物细胞壁特有化合物的合成。我们想看看这种独特类型的酶的机制。3个数据集-它们已被冻结，并在我们的旋转阳极上进行了预筛选，并为旅行存储。还有其他几个类似的突变体正在结晶。如果我们能得到晶体，对它们进行预冷冻和预测试，如果有时间，我们会带着它们来收集。------------------------------------------------------------------我们名单上的每一块晶体都在我们旋转的阳极上进行了预冷冻和预筛选。我们会带一些常温盘子，但纯粹是作为备用。Vrar上的MAD数据是我们此行的首要任务。