Acquisition of an Optima Analytical Ultracentrifuge

购买 Optima 分析超速离心机

• 批准号: 10177290
• 负责人: Edwin Antony
• 金额: $ 51.47万
• 依托单位: SAINT LOUIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-06-01 至 2022-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10177290
• 关键词: 15 year old; Advisory Committees; Age; Arts; Basic Science; Biochemical; Biochemistry; Biophysics; Blood coagulation; Complex; Cryoelectron Microscopy; DNA Repair; Data; Data Analyses; Detection; Development; Fluorescence; Funding; Genetic Recombination; Grant; Heterogeneity; Measurement; Measures; Mechanics; Minor; Modeling; Molecular Biology; Nerve Degeneration; Optics; Postdoctoral Fellow; Preparation; Process; Property; Proteins; Regulation; Research; Research Personnel; Resolution; Saints; Sampling; Schedule; Science; Services; Shapes; Structure; System; Techniques; Time; Training; Translations; Ubiquitination; United States National Institutes of Health; Universities; college; experience; graduate student; immunothrombosis; instrument; instrumentation; interest; medical schools; member; operation; protein aggregation; undergraduate student

ABSTRACT We request funds to purchase an Optima Analytical Ultra Centrifuge (AUC) equipped with interference optics from Beckman Inc. AUC is a powerful, first principle, technique that provides in solution measurement of sample shape, size, and heterogeneity, in addition to measuring strengths of bio-molecular interactions. AUC enables the researcher to perform native-condition characterization of their protein of interest. The instrument will be housed in the Department of Biochemistry and Molecular Biology at the Saint Louis University School of Medicine. The instrument will support basic research of NIH-funded investigators at both the Medical School and in the College of Arts and Sciences. The AUC will also be pivotal in assessing sample homogeneity for structural analysis using cryo-electron microscopy. We describe projects from 4 major users with R01 NIH grants and 4 minor users that will benefit from the requested instrumentation. The user projects span a range of biomolecular interactions in DNA repair and recombination, blood clotting processes, protein aggregation in neurodegeneration, immunothrombosis, regulation of protein translation, laminopathies and ubiquitination processes. All these projects depend on biochemical analysis of the respective protein-protein and protein-substrate complexes and the AUC will be instrumental in establishing their oligomeric and biomolecular interaction properties. The new optima AUC will replace the existing older model ProteomeLab AUC which is now 15 years old and well past the serviceable age. The optima AUC is also redesigned to house the mechanical parts within the main chamber and possess much greater optical resolution and data capture properties. More importantly, the optima AUC is compatible with fluorescence detection capabilities that are currently under development. Training for the optima AUC will be primarily handled by members of the Antony research group who have more than a decade of experience using the instrumentation, sample preparation, and data analysis. Users (undergraduate & graduate students, postdoctoral fellows, and research associates) will be trained to independently use the AUC and instrument time will be determined using the existing scheduling system for departmental instrumentation. A local advisory committee will provide oversight of the AUC and its operations. The optima AUC will be an integral component of the biophysical instrumentation portfolio at Saint Louis University and will have a significant impact on the NIH-funded research.

抽象的 我们要求资金购买配备配备的Optima分析超级离心机（AUC） Beckman Inc.的干扰光学AUC是一种强大的，第一原理，提供的技术 除了测量了样品形状，大小和异质性的溶液测量 生物分子相互作用。 AUC使研究人员能够执行天然条件的表征 他们感兴趣的蛋白质。该仪器将安置在生物化学和分子部 圣路易斯大学医学院的生物学。该工具将支持 NIH资助的医学院和艺术与科学学院的调查员。 AUC 使用冷冻电子评估样品均匀性的样品均匀性也将是关键的 显微镜。我们描述了来自R01 NIH赠款和4位未成年人用户的4个主要用户的项目 受益于要求的仪器。用户项目跨越一系列生物分子互动 在DNA修复和重组中，血液凝结过程，神经退行性中的蛋白质聚集， 免疫栓塞，蛋白质翻译的调节，椎板病和泛素化过程。全部 这些项目取决于对各个蛋白质 - 蛋白质和蛋白质 - 基底物的生化分析 复合物和AUC将有助于建立其寡聚和生物分子 相互作用属性。新的Optima AUC将替换现有的旧型号proteomelab auc 现在已经15岁了，远远超过了可服务的年龄。 Optima AUC也被重新设计为 将机械零件放置在主腔室内，并具有更大的光学分辨率和 数据捕获属性。更重要的是，Optima AUC与荧光检测兼容 目前正在开发的功能。最佳AUC的培训将主要处理 由安东尼研究小组的成员拥有十多年的经验 仪器，样品制备和数据分析。用户（本科生和研究生， 博士后研究员和研究协会将接受培训，以独立使用AUC和 仪器时间将使用现有部门的现有调度系统确定 仪器。当地咨询委员会将监督AUC及其运营。这 Optima AUC将是圣路易斯生物物理仪器投资组合的组成部分 大学，将对NIH资助的研究产生重大影响。

项目成果

Fluorescent human RPA to track assembly dynamics on DNA.

荧光人类 RPA 用于追踪 DNA 上的组装动态。

• DOI: 10.1101/2023.11.23.568455
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Kaushik,Vikas;Chadda,Rahul;Kuppa,Sahiti;Pokhrel,Nilisha;Vayyeti,Abhinav;Grady,Scott;Arnatt,Chris;Antony,Edwin
• 通讯作者: Antony,Edwin

Wrapping of single-stranded DNA by Replication Protein A and modulation through phosphorylation.

复制蛋白 A 包裹单链 DNA 并通过磷酸化进行调节。

• DOI: 10.1101/2024.03.28.587234
• 发表时间: 2024
• 期刊: bioRxiv : the preprint server for biology
• 作者: Chadda,Rahul;Kaushik,Vikas;Ahmad,IramMunir;Deveryshetty,Jaigeeth;Holehouse,Alex;Sigurdsson,SnorriThD;Bothner,Brian;Dastvan,Reza;Origanti,Sofia;Antony,Edwin
• 通讯作者: Antony,Edwin

Mechanistic insight into AP-endonuclease 1 cleavage of abasic sites at stalled replication fork mimics.

• DOI: 10.1093/nar/gkad481
• 发表时间: 2023-07-21
• 期刊: Nucleic acids research
• 影响因子: 14.9

Generation of site-specifically labelled fluorescent human XPA to investigate DNA binding dynamics during nucleotide excision repair.

生成位点特异性标记的荧光人 XPA，用于研究核苷酸切除修复过程中的 DNA 结合动力学。

• DOI: 10.1016/j.ymeth.2024.02.006
• 发表时间: 2024
• 期刊: Methods (San Diego, Calif.)
• 作者: Kuppa,Sahiti;Corless,Elliot;Caldwell,ColleenC;Spies,Maria;Antony,Edwin
• 通讯作者: Antony,Edwin

Interplay of macromolecular interactions during assembly of human DNA polymerase δ holoenzymes and initiation of DNA synthesis.

人类 DNA 聚合酶 δ 全酶组装和 DNA 合成启动过程中大分子相互作用的相互作用。

• DOI: 10.1101/2023.05.09.539896
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Norris,JessicaL;Rogers,LindseyO;Pytko,KaraG;Dannenberg,RachelL;Perreault,Samuel;Kaushik,Vikas;Kuppa,Sahiti;Antony,Edwin;Hedglin,Mark
• 通讯作者: Hedglin,Mark