BIOPHYSICAL CHARACTERIZATION OF MACROMOLECULES

大分子的生物物理表征

• 批准号: 6290696
• 负责人: PETER SCHUCK
• 金额: --
• 依托单位: OFFICE OF THE DIRECTOR, NATIONAL INSTITUTES OF HEALTH
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6290696
• 关键词: antibody; antibody receptor; biological transport; biomedical equipment development; biosensor device; chemical binding; chemical kinetics; fluidity; immunoglobulin E; intermolecular interaction; lysozyme; method development; surface plasmon resonance; thermodynamics

The intent of this project is to develop new and improve existing methodology for the characterization of biological macromolecules, and to apply these methods collaboratively to the study of macromolecules and their interactions. Techniques employed are analytical ultracentrifugation, dynamic light scattering, isothermal titration calorimetry, and surface plasmon resonance biosensing. In analytical ultracentrifugation, substantial methodological advances have been made in the modeling of sedimentation velocity data by direct boundary analysis. This includes a novel algebraic noise decomposition method, a least-squares g*(s) method, and the analysis of continuous size distributions. Additional experience was gained in analytical zone centrifugation, a sedimentation velocity method that minimizes the required sample volume. Also, we continued the development of methods for the hydrodynamic characterization of small molecules. We have expanded our expertise in dynamic light scattering and isothermal titration calorimetry, and have written a review on the experimental methods applied in optical biosensing. Experimentally, one major focus of the systems collaboratively studied has been the interactions of membrane receptors and related proteins important in immunology, such as the T-cell receptor, different NK receptors, MHC molecules, superantigen, and FcRn-receptor. A second emphasis was in viral proteins, including HIV-gp120 and herpes simplex capsid proteins. Further systems studied are G-protein subunit interactions, calmodulin interactions with neurogranin, as well as nucleosomes and their interaction with HMG proteins. The characterization of many of these systems has been completed, and several collaborative publications are in press, submitted, and in preparation. - biophysics, binding, thermodynamics, kinetics

这个项目的目的是开发新的和改进现有的表征生物大分子的方法，并将这些方法协同应用于大分子及其相互作用的研究。采用的技术有分析超速离心法、动态光散射技术、等温滴定量热法和表面等离子体共振生物传感技术。在分析超速离心法中，直接边界分析在模拟沉积速度数据方面取得了很大的方法学进展。这包括一种新的代数噪声分解方法，一种最小二乘g*(S)方法，以及连续尺寸分布的分析。在分析区离心法方面获得了其他经验，这是一种将所需样本量降至最低的沉降速度法。此外，我们还继续开发小分子的流体动力学表征方法。我们扩展了我们在动态光散射和等温滴定量热法方面的专业知识，并对应用于光学生物传感的实验方法进行了综述。在实验上，共同研究的系统的一个主要焦点是膜受体与免疫学中重要的相关蛋白的相互作用，如T细胞受体、不同的NK受体、MHC分子、超抗原和FcRN受体。第二个重点是病毒蛋白，包括HIV-gp120和单纯疱疹病毒衣壳蛋白。进一步研究的系统包括G蛋白亚基相互作用，钙调蛋白与神经颗粒素的相互作用，以及核小体及其与HMG蛋白的相互作用。其中许多系统的特点已经完成，几份合作出版物正在出版、提交和筹备中。-生物物理学、结合、热力学、动力学