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

该项目的目的是开发新的和改善现有的方法来表征生物大分子的表征，并将这些方法合作应用于大分子及其相互作用的研究。所采用的技术是分析性超速离心，动态光散射，等温滴定量热法和表面等离子体共振生物传感。在分析性超速离心中，通过直接边界分析在沉积速度数据建模中取得了实质性的方法学进步。这包括一种新型的代数噪声分解方法，最小二乘G*（S）方法以及连续大小分布的分析。在分析区离心区中获得了额外的经验，这是一种减少所需样品体积的沉积速度方法。此外，我们继续开发小分子流体动力学表征的方法。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，并就光学生物传感应用中应用的实验方法进行了评论。在实验上，对系统进行了协作的主要重点是膜受体和相关蛋白在免疫学中重要的蛋白质的相互作用，例如T细胞受体，不同的NK受体，MHC分子，超级抗原和FCRN受体。第二个重点是病毒蛋白，包括HIV-GP120和疱疹单纯胶囊蛋白。研究的进一步系统是G蛋白亚基相互作用，钙调蛋白与神经素蛋白以及核小体的相互作用及其与HMG蛋白的相互作用。这些系统的表征已经完成，并且在媒体上，提交并准备了一些协作出版物。 - 生物物理学，结合，热力学，动力学