Introduction of Fluorometry to the Screening of Protein Crystallization Buffers

Abstract

A system to use fluorometry for the search of protein crystallization buffers was developed. The screening of candidates was done with a use of short gel-filtration column (Sephacryl S-100 HR) linked to the fluorometer. Protein elution was monitored by intrinsic fluorescence or emission from its labels. This method was applied to actin and actin complexes. When nuclei were formed in actin solution, preceding the peak of actin, a new peak appeared on the elution curve. It was found that the fluorescence allows detection of even small amount of nuclei formed in the buffer. Using the screened buffers, crystal growths were attempted. The images of crystals were captured by fluorescence microscope. The monitoring of nuclei with this method will accelerate the screening of crystallization buffers. This system is applicable to the crystallization of other proteins and their complexes.