Development of the concept for structure-aided rational design of bright fluorescent protein

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Fluorescent proteins are used as indispensable tool for biological research. However, enhancement of the fluorescence intensity has been accomplished by repeating the random mutagenesis, thus the effective way for improvement has been required. In this study we determined structures of the intermediates those were generated through the improvement of the fluorescence intensity for a bright fluorescent protein. From the comparison among them, we found structural features contributing to the improvement. In the mutants fluorescence intensity has enhanced, planarity of fluorescent chromophore is enhanced and main chain of the beta strand comes close to the fluorescent chromophore for stabilization. In combination with currently creating molecular modeling system based on the computational simulation, establishment of the rational design concept for bright fluorescent protein is expected by further study.