ATOMIC FORCE MICROSCOPY OF BIOPOLYMERS

生物聚合物的原子力显微镜

• 批准号: 8365490
• 负责人: Catherine E. Costello
• 金额: $ 1.85万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-06-01 至 2012-08-09
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8365490
• 关键词: Amino Acid Sequence; Amyloid Fibrils; Amyloid Proteins; Amyloidosis; Atomic Force Microscopy; Biology; Biophysics; Biopolymers; Boston; Calibration; Carbon Nanotubes; Collagen; Complement; Complex; Development; Devices; Doctor of Philosophy; Funding; GAG Gene; Goals; Grant; Image; Individual; Maintenance; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Medicine; Molecular; Molecular Weight; Monitor; Mucins; National Center for Research Resources; Organ; Patients; Post-Translational Protein Processing; Principal Investigator; Property; Proteins; Proteoglycan; Recombinants; Research; Research Infrastructure; Resolution; Resources; Sampling; Source; Supervision; Testing; Tissues; Training; Tube; United States National Institutes of Health; Universities; Variant; Weight; amyloid fibril formation; base; cost; glycosylation; improved; instrument; operation; single molecule; 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 BUSM MS Resource carries out atomic force microscopy to complement mass spectrometry studies of individual biopolymers of high molecular weight, including, e.g., proteoglycans, collagens and molecules having collagen-type domains, and other proteins that contain high levels of glycosylation, as well as non-covalent complexes. AFM is being used to estimate the overall weight and modular composition of such species, on the basis of extrapolations of calibration curves over the range accessible by mass spectrometry. The aim of this project is to use AFM to quickly estimate the MWs of large molecules and complexes as a supplemental tool for mass spectrometry in the high molecular weight range. This is achieved by using AFM to measure the molecular volume of single molecules of known MW as standards, extrapolating a curve using biopolymer standards at different MW, and determining the relation between molecular volume and MW. AFM is also being used to characterize amyloid fibrils, with the goal of relating fibril properties to variations in the amino acid sequence and to posttranslational modifications of the constituent proteins. By using purified patient samples as well as recombinant amyloid proteins, the formation of amyloid fibrils under different conditions and protein/GAG interactions is monitored in both this development project and in several collaborative projects (e.g., Nugent, Skinner, Trinkaus-Randall). AFM images have also been recorded for amyloid fibrils obtained from organ tissues of patients who have succumbed to primary amyloid disease. A manuscript describing recent results will be submitted shortly. To improve resolution in AFM measurements, the use of carbon nanotubes is being explored and a device for mounting the tubes has been constructed and tested. Day-to-day operation, maintenance, training and supervision of the instrument are the responsibility of Dr. Hong, who completed his PhD in the Boston University Dept. of Cellular Biophysics with a focus on AFM studies of mucins.

该子项目是利用资源的许多研究子项目之一 由NIH/NCRR资助的中心赠款提供。次级项目的主要支助 子项目的主要研究者可能是由其他来源提供的， 包括其他NIH来源。 为子项目列出的总成本可能 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。 BUSM MS资源进行原子力显微镜，以补充高分子量的单个生物聚合物的质谱研究，包括，例如，蛋白聚糖、胶原和具有胶原型结构域的分子，以及含有高水平糖基化的其它蛋白质，以及非共价复合物。原子力显微镜正被用来估计这些物种的总重量和模块组成的基础上，外推的校准曲线的范围内可通过质谱仪。本项目的目的是利用原子力显微镜快速估计大分子和复合物的分子量，作为质谱在高分子量范围内的补充工具。这是通过使用AFM测量已知MW的单分子的分子体积作为标准，使用不同MW的生物聚合物标准外推曲线，并确定分子体积和MW之间的关系来实现的。原子力显微镜也被用来表征淀粉样蛋白原纤维，与原纤维的性质有关的氨基酸序列的变化和翻译后修饰的组成蛋白质的目标。 通过使用纯化的患者样品以及重组淀粉样蛋白，在该开发项目和几个合作项目（例如，Nugent，Skinner，Trinkaus-Randall）.从原发性淀粉样蛋白病患者的器官组织中获得的淀粉样蛋白原纤维也记录了AFM图像。不久将提交一份介绍最近成果的手稿。为了提高原子力显微镜测量的分辨率，正在探索使用碳纳米管和安装管的设备已经构建和测试。 仪器的日常操作、维护、培训和监督由洪博士负责，他在波士顿大学医学系完成了博士学位。细胞生物物理学的重点是粘蛋白的AFM研究。