NOVEL CONTINUOUS FLOW MIXER DESIGN FOR TIME RESOLVED VISIBLE & IR STUDIES

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。在理解蛋白质折叠动力学方面的最新进展与利用光学触发或改进的快速混合方法在微秒时间尺度上引发和观察构象变化的新实验方法的发展密切相关。虽然在1923年首次报道了快速反应的连续流测量，但由于消耗的样品量很大，该方法的后续使用有限。我们最近的研究表明，通过使用一组探测器同时监测流道沿线大量点的信号，可以大大提高该方法的效率。通过将这种灵敏的检测方法与高效的毛细管混合器相结合，我们能够实现死区时间短至45 us。该项目的目标是将这种改进的快速混合技术与配备红外敏感探测器阵列的傅里叶变换红外光谱仪相结合，以实时监测红外波长下的蛋白质折叠事件。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Recent progress in understanding the dynamics of protein folding has been closely linked with the development of new experimental approaches for initating and observing conformational changes on the microsecond time scale, using optical triggers or improved rapid mixing methods. Although the first continuous-flow measurement of a rapid reaction has been reported in 1923, the method has enjoyed only limited subsequent use because of the large amount of sample consumed. We have recently shown that the efficiency of the method can be greatly improved by using an array of detectors to simultaneously monitor the signal at a large number of points along the flow channel. By combining this sensitive detection method with an efficient capillary mixer, we were able to achieve dead times as short as 45 us. The goal of this project is to combine this improved rapid mixing technique with a Fourier-Transform IR spectrometer equipped with an IR sensitive detector array to monitor protein folding events in real time at infrared wavelengths.