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

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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.