A Stroboscopic Time-Resolved Spectrofluorometer

描述(由申请人提供)：七名在蛋白质相互作用为主要组成部分的领域工作的BRI科学家提议购买一台闪光时间分辨荧光光谱仪，该仪器将能够测量从纳秒到秒的荧光和磷光寿命、各向异性衰减和稳态光子计数光谱测量。该仪器将能够研究蛋白质结构和相互作用的内在和外在探针的发射，并提供关于探针所在区域的可及性、环境和灵活性的信息。此外，还将能够通过FRET和LRET(分别为Forster和荧光共振能量转移)测量距离、距离分布和距离变化。结合谱的灵敏测量将允许获得结合常数和化学计量以及荧光参数的热依赖关系。 该仪器将促进研究人员目前的实验方法，包括X射线结晶学、荧光和磷光光谱、差示扫描和等温滴定热法、圆二色谱、质谱学和其他生化/生物物理测量。时间分辨光谱仪将补充NIH资助的结构和功能研究，重点是了解平滑肌、心肌和骨骼肌的调节、伴侣的功能、参与骨骼肌和心肌兴奋收缩偶联的蛋白质以及参与炭疽感染的蛋白质。这些研究将为了解包括心脏和骨骼肌病、高血压、动脉粥样硬化和病毒感染在内的疾病状态提供基础。

DESCRIPTION (provided by applicant): Seven BBRI scientists working in fields in which protein interactions are a primary component, propose to purchase a Stroboscopic Time-Resolved Spectrofluorometer which will enable fluorescence and phosphorescence lifetime measurements from nanoseconds to seconds, anisotropy decay and steady-state photon counting spectral measurements. The instrument will enable study of the emission of intrinsic and extrinsic probes of protein structure and interactions and provide information about the accessibility, environment, and flexibility of the domain to which the probe is attached. In addition, measurements of distance, distance distributions, and distance changes via FRET and LRET (Forster and fluorescence resonance energy transfer, respectively) will be enabled. Sensitive measurements of binding profiles will allow binding constants and stoichiometry and thermal dependence of fluorescence parameters to be obtained. This instrument will facilitate the current experimental approaches of the investigators which include, X-ray crystallography, fluorescence and phosphorescence spectroscopy, differential scanning and isothermal titration calorimetry, circular dichroism, mass spectroscopy and other biochemical / biophysical measurements. The time-resolved spectrofluometer will complement NIH funded structural and functional studies focused on understanding the regulation of smooth, cardiac and skeletal muscle, the function of chaperones, proteins involved in excitation-contraction coupling in skeletal and cardiac muscle and proteins involved in anthrax infection. These studies will provide the basis for understanding the diseased state including cardiac and skeletal myopathies, hypertension, atherosclerosis, and viral infection.