BIOPHYSICAL EVALUATION OF SOL-GEL ENCAPSULATED PROTEINS

溶胶-凝胶封装蛋白质的生物物理评价

• 批准号: 6530743
• 负责人: JOEL M FRIEDMAN
• 金额: $ 33.21万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2004-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6530743
• 关键词: Raman spectrometry; biophysics; chemical models; conformation; fluorescence spectrometry; gel; hemoprotein; mathematical model; physical state; spectrometry; structural biology

DESCRIPTION: (applicant's abstract) The encapsulation of proteins in porous sol-gels has captured the attention of both applied and basic research scientists because the encapsulated proteins retain functional activity and exhibit enhanced stability. Despite the widespread interest in using this technology to develop new classes of robust biosensors, no in-depth biophysical study of protein response to sol-gel encapsulation has been carried out. The project proposed herein will systematically examine the impact of three promising encapsulation protocols on the conformation and dynamics of the well-characterized heme proteins, hemoglobin and myoglobin. We have probed the solution-phase conformation - secondary through quaternary - of these heme proteins with a myriad of laser-based time-resolved and steady state spectroscopic techniques, and are therefore uniquely qualified to examine the dynamic and static heme protein/sol-gel interactions. The spectroscopic tools at our fingertips include: CW UV resonance Raman, CW and time-resolved visible resonance Raman, time-resolved near IR absorption, time resolved fluorescence anisotropy and fluorescence lifetime measurements. Subtle as well as large-scale conformational changes and the associated dynamics are probed when combining these techniques. In addition, nanosecond, time-resolved absorption spectroscopy will primarily be used to characterize the ligand rebinding kinetics of the heme proteins in the sol-gels. It is anticipated that this comprehensive study will result in an algorithm for matching an encapsulation protocol to a particular biomedical or biophysical application.

描述：（申请人的摘要）将蛋白质封装在多孔中 溶胶-凝胶引起了应用和基础研究的关注 科学家们因为封装的蛋白质保留了功能活性 表现出增强的稳定性。尽管人们对使用此技术产生了广泛的兴趣 开发新型稳健生物传感器的技术，无需深入的生物物理学 已经进行了蛋白质对溶胶-凝胶封装反应的研究。这 本文提出的项目将系统地研究三个方面的影响 关于构象和动力学的有前途的封装协议 充分表征的血红蛋白、血红蛋白和肌红蛋白。我们已经探究了 这些血红素的溶液相构象 - 二级到四级 具有多种基于激光的时间分辨和稳态的蛋白质 光谱技术，因此具有独特的资格来检查 动态和静态血红素蛋白/溶胶-凝胶相互作用。光谱工具 我们触手可及的包括：连续波紫外共振拉曼、连续波和时间分辨可见光 共振拉曼、时间分辨近红外吸收、时间分辨荧光 各向异性和荧光寿命测量。微妙也 大规模构象变化和相关动力学被探测 结合这些技术。此外，纳秒、时间分辨吸收 光谱将主要用于表征配体重新结合 溶胶-凝胶中血红素蛋白的动力学。预计这 综合研究将产生匹配封装的算法 特定生物医学或生物物理应用的协议。