Correlating spectroscopic properties of fluorescent proteins (FPs) tot heir crystal structure: towards the rational design of FPs for superresolution microscopy.

Superresolution microscopy consists of several techniques, which are often used for the visualization of subcellular systems. Because most of these techniques are based on the specific dynamic behavior of the dynamic fluorescent label, they all have their own advantages and disadvantages and the obtained results will therefore be different. Therefore, the combination of different techniques (multimodal superresolution microscopy) would allow researchers to increase the information content of their experiments and allow them to validate their superresolution results. A good example is the combination of PhotoActivated Localization Microscopy (PALM) and Photochromic Stochastic fluctuation imaging (pcSOFI). However, multimodal suerresolution microscopy demands for specific dynamic fluorescent proteins and introducing the desired dynamic characteristics into fluorescent proteins using random mutagenesis and directed evolution is a costly and time-consuming process. For a more efficient rational design of these dynamic fluorescent proteins, more knowledge about them is necessary. Therefore, the relationship between molecular structureand function of dynamic fluorescent proteins will be investigated. During this project, different cycles of mutagenesis, characterization, crystallization, structure determination and statistical analysis will be performed. Finally, we will be able to design dynamic fluorescent proteinssuitable for multimodal microscopy.

Keywords:Protein design, Protein crystallography, Multimodal superresolution Fluorescence

Disciplines:Biochemistry and metabolism, Medical biochemistry and metabolism, Systems biology

Project type:PhD project