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Time-Resolved Diffusion Detection with Microstopped Flow System
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DOI：10.1021/acs.analchem.9b02897.png)
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Abstract
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The transient grating (TG) method is a powerful technique for monitoring the time dependence of the diffusion coefficient during photochemical reactions. However, the applications of this technique have been limited to photochemical reactions. Here, a microstopped flow (mu-SF) system is developed to expand the technique's applicability. The constructed mu-SF system can be used for a solution with a total volume as small as 3 mu L, and mixing times for absorption and diffusion measurements were determined to be 400 mu s and 100 ms, respectively. To demonstrate this system with the TG method, an acid-induced denaturation of a photosensor protein, phototropin LOV2 domain with a linker, was studied from the viewpoint of the reactivity. This system can be used not only for time-resolved diffusion measurement but also for conventional absorption or fluorescence detection methods. In particular, this system has a great advantage for a target solution in that only a very small amount is needed.
Keywords:
INTERMOLECULAR INTERACTION
ARABIDOPSIS PHOTOTROPIN-1
DYNAMICS
LOV2
BINDING
DOMAIN
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