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

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。所列机构为中心机构，不一定是研究者机构。最近在理解蛋白质折叠动力学方面的进展与新的实验方法的发展密切相关，这些实验方法用于在微秒时间尺度上启动和观察构象变化，使用光学触发器或改进的快速混合方法。虽然第一个连续流动测量快速反应的方法在1923年就有报道，但由于消耗了大量的样品，该方法随后的应用非常有限。我们最近已经表明，该方法的效率可以大大提高，通过使用一个阵列的检测器，同时监测信号在大量的点沿着流动通道。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，我们能够实现短至45 us的死区时间。该项目的目标是将这种改进的快速混合技术与配备有红外敏感检测器阵列的傅里叶变换红外光谱仪联合收割机相结合，以在红外波长下真实的时间监测蛋白质折叠事件。

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.