ATOMIC FORCE MICROSCOPY OF BIOPOLYMERS

生物聚合物的原子力显微镜

• 批准号: 8170853
• 负责人: Catherine E. Costello
• 金额: $ 2.33万
• 依托单位: BOSTON UNIVERSITY MEDICAL CAMPUS
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-06-01 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8170853
• 关键词: Amino Acid Sequence; Amyloid Fibrils; Amyloid Proteins; Amyloid beta-Protein; Amyloidosis; Atomic Force Microscopy; Biophysics; Biopolymers; Boston; Calibration; Carbon Nanotubes; Collagen; Complement; Complex; Computer Retrieval of Information on Scientific Projects Database; Development; Devices; Doctor of Philosophy; Electronic Mail; Funding; Goals; Grant; Image; Individual; Institution; Maintenance; Manuscripts; Mass Spectrum Analysis; Measurement; Measures; Molecular; Molecular Weight; Monitor; Mucins; North Carolina; Organ; Patients; Post-Translational Protein Processing; Property; Proteins; Proteoglycan; Recombinants; Research; Research Personnel; Resolution; Resources; Sampling; Source; Supervision; Telephone; Testing; Tissues; Tube; United States National Institutes of Health; Universities; Variant; Weight; amyloid fibril formation; base; experience; glycosylation; improved; instrument; macromolecule; operation; single molecule; synuclein; tool

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. 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 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. Dr. Ding, who has extensive experience in AFM of beta-amyloid and synuclein amyloid fibrils and various types of macromolecules, helped in the planning for this project and continues to provide advice and assistance to the BUSM investigators. Although he has relocated to North Carolina, he remains in email and phone contact.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 BUSM MS资源进行了原子力显微镜，以补充高分子重量的个体生物聚合物的质谱研究，包括，例如，蛋白聚糖，胶原蛋白和具有胶原蛋白型结构域的蛋白聚糖，胶原蛋白和分子，以及其他含有高水平的糖基化水平的蛋白质，以及非胆汁糖基化的复合物。根据质谱法可以访问的范围内校准曲线的外推，AFM用于估计此类物种的总体重量和模块化组成。该项目的目的是使用AFM快速估计大分子和复合物的MW作为高分子重量范围内质谱法的补充工具。这是通过使用AFM来测量已知MW标准品的单个分子的分子体积，使用不同MW处的生物聚合物标准来推断曲线的分子体积，并确定分子体积和MW之间的关系。 AFM还用于表征淀粉样蛋白原纤维，其目的是将原纤维性质与氨基酸序列的变化和成分蛋白的翻译后修饰有关。 通过使用纯化的患者样品以及重组淀粉样蛋白，在该开发项目和多个协作项目中都可以监测淀粉样蛋白原纤维在不同条件和蛋白质/GAG相互作用下的形成（例如Nugent，Skinner，Trinkaus-randall）。还记录了从屈服于原发性淀粉样蛋白疾病的患者的器官组织中获得的淀粉样蛋白原纤维的记录。描述最新结果的手稿将很快提交。为了改善AFM测量的分辨率，正在探索碳纳米管的使用，并构建和测试了用于安装管的装置。 该工具的日常运作，维护和监督是Hong博士的责任，他在波士顿大学的蜂窝生物物理学部完成了博士学位，重点是对粘蛋白的AFM研究。 Ding博士在AFM拥有β-淀粉样蛋白和突触核蛋白淀粉样蛋白原纤维以及各种类型的大分子的AFM经验，他为该项目的计划提供了帮助，并继续为Busm的调查人员提供建议和帮助。 尽管他已搬到北卡罗来纳州，但他仍在电子邮件和电话联系中。