kT-scale Protein & Carbohydrate Interactions on Synthetic Materials & Cell Surfaces

kT级蛋白质

• 批准号: 1402739
• 负责人: Michael Bevan
• 金额: $ 30万
• 依托单位: Johns Hopkins University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-06-01 至 2018-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1402739&HistoricalAwards=false
• 关键词: kT; scale; Protein; & Carbohydrate

PI: Bevan, Michael A. Proposal Number: 1402739Institution: Johns Hopkins University Title: kT-scale Protein & Carbohydrate Interactions on Synthetic Materials & Cell Surfaces Many important cellular interactions involve "weak" interactions, which are difficult to characterize by current techniques. The PI proposes to move into new territory, in terms of directly characterizing these weak intermolecular interactions of biomolecules, by using a technique, Diffusing Colloidal Probe Microscopy (DCPM), developed by the PI's group. The proposal will examine two systems of biological importance, which were also chosen for their significance to biomedical problems: stem-cell based tissue engineering and cancer metastasis. Success in the development of the technique for studying weak biological interactions will have an impact on the understanding of such interactions in cell-cell adhesion, cell migration, tissue morphogenesis, metastasis, immunology, and other cellular processes.Weak biomolecular interactions, on the order of kT, play an important role in cell-cell adhesion, cell migration, tissue morphogenesis, metastasis, immunology, and other cellular processes. While there are standard mechanical methods for measuring strong interactions (pN forces), weak interactions are generally measured_ indirectly_ by spectroscopic means. The PI proposes to move into new territory, by directly characterizing these interactions using DCPM, which offers spatial and temporal resolution, statistical significance, and directness not accessible using scanning probes or spectroscopic methods. In essence, DCPM passively monitors Brownian excursions of "diffusing probes" and exploits these natural gauges for time (a2/D), energy (kT), force (fN), and length (nm) when interrogating the interaction of proteins and carbohydrates. This is inherently a nanoscale technique because it harnesses Brownian motion as a useful tool to directly measure kT interactions between proteins rather than avoiding stochastic thermal motion as an undesirable complication of nanoscale systems. Two systems will be studied, CD44-HA interactions and heterophilic cadherin interactions (in the presence of calcium ions) on supported bilayers. The development of DCPM for studying weak biological interactions will have an impact on the understanding of such interactions in cell-cell adhesion, cell migration, tissue morphogenesis, metastasis, immunology, and other cellular processes.

PI：Bevan, Michael A. 提案编号：1402739 机构：约翰霍普金斯大学 标题：合成材料和细胞表面上的 kT 尺度蛋白质和碳水化合物相互作用 许多重要的细胞相互作用涉及“弱”相互作用，目前的技术很难表征这些相互作用。 PI 提议进入新领域，通过使用 PI 小组开发的扩散胶体探针显微镜 (DCPM) 技术，直接表征生物分子的这些弱分子间相互作用。该提案将研究两个具有生物学重要性的系统，这两个系统也因其对生物医学问题的重要性而被选中：基于干细胞的组织工程和癌症转移。研究弱生物相互作用的技术开发的成功将对细胞-细胞粘附、细胞迁移、组织形态发生、转移、免疫学和其他细胞过程中的相互作用的理解产生影响。kT量级的弱生物分子相互作用在细胞-细胞粘附、细胞迁移、组织形态发生、转移、免疫学和其他细胞过程中发挥重要作用。虽然有测量强相互作用（pN 力）的标准机械方法，但弱相互作用通常是通过光谱手段间接测量的。 PI 提议进入新领域，通过使用 DCPM 直接表征这些相互作用，DCPM 提供空间和时间分辨率、统计显着性以及使用扫描探针或光谱方法无法获得的直接性。本质上，DCPM 被动地监测“扩散探针”的布朗偏移，并在询问蛋白质和碳水化合物的相互作用时利用这些自然量规来测量时间 (a2/D)、能量 (kT)、力 (fN) 和长度 (nm)。这本质上是一种纳米级技术，因为它利用布朗运动作为直接测量蛋白质之间 kT 相互作用的有用工具，而不是避免随机热运动作为纳米级系统的不良并发症。将研究两个系统：CD44-HA 相互作用和支持双层上的异嗜性钙粘蛋白相互作用（在钙离子存在下）。用于研究弱生物相互作用的 DCPM 的发展将影响对细胞间粘附、细胞迁移、组织形态发生、转移、免疫学和其他细胞过程中此类相互作用的理解。