Optimization of red fluorescent marker proteins guided by evaluation of structure-function relations
FacultiesFakultät für Naturwissenschaften
LicenseStandard (Fassung vom 01.10.2008)
Red fluorescent proteins (RFPs) attract particular attention from both scientific and practical points of view. They have a great potential to be used as in vivo markers in multicolour labelling and FRET-based applications. Since long-wave light penetrates tissue more easily, far-red FPs provide an excellent tool for whole-body imaging. Besides the discovery of naturally occurring RFPs, they can also be generated by mutagenesis of non-fluorescent chromoproteins. However, the key residues responsible for a bright red FP have to be identified. Therefore, we investigated both non-fluorescent and high fluorescent GFP-like proteins from Anemonia sulcata var. rufescens. In the course of further development of fluorescent proteins, a new class of probes opened up advances in dynamic cellular imaging. These optical highlighters are able to modify their fluorescence emission colour and intensity, respectively, due to wavelength-dependent illumination. The reversible on- and off-switching between a fluorescent and a non-fluorescent chromophore state is characteristic for a specific class of optical highlighters. A cis-trans isomerization of the chromophore was revealed as key event in the switching process in recent studies concerning the molecular switching mechanism of photoswitchable fluorescent proteins (PSFPs). A promising area of application for PSFPs is their usage in high resolution fluorescence microscopy. Due to the increasing demand for novel PSFPs, we investigated various red FPs from Anthozoa to obtain a new member of this class of optical highlighters. The widespread distribution of GFP-like proteins in zooxanthellate and azooxanthellate cnidarians and their impressive colour diversity indicates different specific biological functions, which are still not completely understood. For this reason, field studies concerning two different Anemonia species in combination with investigation of host pigmentation and several biomarkers were performed during this thesis.
Subject HeadingsFluoreszenzmarkierung [GND]