Time-Resolved FTIR System for Biological Applications

用于生物应用的时间分辨 FTIR 系统

• 批准号: 9513715
• 负责人: Kenneth Rothschild
• 金额: --
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-01 至 1999-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9513715&HistoricalAwards=false
• 关键词: Time; Resolved; FTIR; System; Biological

The central goal of this project is the development of advanced time-resolved FTIR instrumentation which will allow for the first time the investigation of a wide variety of biological systems at the sub-nanosecond to millisecond range. The proposed instrument will incorporate novel features which will address the major short-comings of current time-resolved FTIR instruments. The project will be supported by the newly formed Center for Photonics Research at Boston University and Bruker Instruments, Inc., a leader in time-resolved FTIR instrumentation In recent years it has been demonstrated that Fourier transform infrared (FTIR) difference spectroscopy can provide detailed information about dynamic changes in the protonation state, local environment and orientation of specific amino acid residues in proteins. This ability is enhanced by the recent development of powerful new methods for band assignments by the introduction of site-directed isotope labels (SDIL) into proteins. However, the current generation of FTIR instruments have fallen short of their promise for probing rapid conformational changes in biomolecular systems. Existing limitations are related to the lack of sufficiently powerful broad-band infrared sources, the relatively slow cycle-time or irreversible response of most biological systems upon activation, the prevalence of biological systems activated by mechanisms other than light, and the need to simultaneously monitor changes in the infrared and UV-visible absorption of a sample. We propose to develop a unique step-scan FTIR system with the ability to record simultaneous sub-nanosecond to millisecond infrared and W-visible absorption changes in biomolecular systems. The instrument will be based on recent breakthroughs in technology for ultra-high intensity broad-band continuous (CW) and pulsed IR sources, rapid sub-nanosecond transient recorders and infrared detectors and step-scan interferometers. The development of a novel synchronized sample step translator (SSST) will make it possible to shorten dramatically measurement times and open the door to the investigation of dynamic processes in a wide variety of biomolecular systems under physiological conditions, including systems not activated by light, systems with very slow cycle times and systems with irreversible processes. By combining this proposed FTIR instrumentation with the recently developed method of SDIL and site-directed non-native amino acid replacement (SNAAR), breakthroughs are expected in the fields of protein folding, enzyme mechanism, ion transport and signal reception. Transfer of the developed technology for future commercial use will be facilitated by the close interaction during the project with Bruker Instruments and through the Center for Photonics Research

该项目的中心目标是开发先进的时间分辨FTIR仪器，这将首次允许在亚纳秒到毫秒范围内对各种生物系统进行调查。拟议的仪器将纳入新的功能，这将解决目前的时间分辨傅里叶变换红外光谱仪的主要缺点。该项目将得到波士顿大学新成立的光子学研究中心和布鲁克仪器公司的支持，时间分辨FTIR仪器的领导者 近年来，傅里叶变换红外（FTIR）差谱技术被证明可以提供蛋白质中特定氨基酸残基的质子化状态、局部环境和取向等动态变化的详细信息。这种能力是增强了最近开发的强大的新方法，通过引入定点同位素标记（SDIL）到蛋白质的频带分配。然而，目前这一代的傅里叶变换红外光谱仪已经达不到他们的承诺，探测快速构象变化的生物分子系统。现有的局限性与缺乏足够强大的宽带红外源，相对缓慢的周期时间或不可逆的反应，大多数生物系统激活后，由机制激活的生物系统的流行，而不是光，并需要同时监测样品的红外和紫外-可见吸收的变化。 我们建议开发一种独特的步进扫描FTIR系统，能够同时记录生物分子系统中的亚纳秒到毫秒的红外和W-可见光吸收变化。该仪器将以超高强度宽带连续和脉冲红外光源、快速亚纳秒瞬态记录器和红外探测器以及步进扫描干涉仪等技术的最新突破为基础。一种新型的同步样品步翻译器（SSST）的发展将有可能显着缩短测量时间，并打开大门，在生理条件下，在各种生物分子系统，包括系统不被光激活，系统具有非常慢的周期时间和系统的不可逆过程的动态过程的调查。通过将这种FTIR仪器与最近开发的SDIL和定点非天然氨基酸置换（SNAAR）方法相结合，有望在蛋白质折叠、酶机制、离子转运和信号接收等领域取得突破。 在项目期间与布鲁克仪器公司的密切互动以及通过光子学研究中心，将促进开发的技术转让用于未来的商业用途