A xanthene-BODIPY cassette is used as a ratiometric intracellular pH reporter for imaging protein-dye conjugates in living cells. A model was hypothesized to explain the pH-dependent energy transfer efficiencies from the donor to the acceptor based on the electronic chemistry data. Sulfonation conditions were developed for BODIPY dyes to give water-soluble functionalized monosulfonation and disulfonation donors. A water-soluble TBET cassette, which has good photophysical properties, was synthesized using a bissulfonated BODIPY dye as the donor, and their applications for in vitro protein labeling is achieved. Chemoselective cross-coupling reactions were demonstrated for C-S bonds in the BODIPY dye, and similar reactions were applied to make the acceptor of the watersoluble cassette. Chemiluminescent energy transfer cassettes based on fluorescein and Nile Red were synthesized and their spectral properties were studied. Pep-1 (also known as Chariot), R8 (which is not often used as a non-covalent protein carrier), and a new synthesized compound, Azo-R8, was used for the study of non-covalent delivery of four different proteins into mammalian cells. Data from confocal spectroscopy revealed that all three carriers are effective for translocating protein cargos into live cells. At 37 dgrees C, import into endocytic compartments dominates, but at 4 degrees C weak, diffuse fluorescence is observed in the cytosol indicative of a favorable mode of action.

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In der vorliegenden Dissertation konnten erfolgreich neben der Entwicklung und Optimierung neuer Methoden in der Kettenverlängerung von Aldosen und Ketosen auch neue Farbstoffe zur Untersuchung von Protonierungs- und Wasserdynamiken designt und synthetisiert werden. Die Erweiterung des Verständnisses im Zusammenspiel von Aminosäuren und Kohlenhydraten hat einen großen Einfluss in der wissenschaftlichen Gesellschaft der Kohlenhydratchemie. Die Kontrolle im Aufbau von Stereotetraden und -pentaden führt zu einer Ausweitung der bekannten Möglichkeiten in der stereoselektiven Synthese von Naturstoffen und Biomimetika. Außerdem konnten durch eine einfache Methode die C-Glykoside seltener Kohlenhydrate dargestellt werden. Einbau von Aminosäuren in Kohlenhydratstrukturen konnten in hohen Stereoselektivitäten und der Möglichkeit der Stereomanipulation durch Wahl des Isocyanides erreicht werden. Die Synthese des zellgängigen Farbstoffes PAc-SNARF, sowie des Biolinkers IA-SNARF ermöglicht eine bildgebenden ratiometrischen pH-Untersuchungen in Zellen und auf Proteinoberflächen. Die Verallgemeinerung der Synthese der Farbstoff-Precursor für die SNARF-Derivate über eine Friedel-Crafts-Acylierung erlaubt eine kostengünstige Darstellung einer großen Bandbreite von Farbstoffen. Mithilfe der neu synthetisierten sterisch anspruchsvollen N-Methyl-6-oxychinoliniumbetain-Derivate verbunden mit der Fluoreszenzaufkonvertierungs-Spektroskopie konnte eine Verlangsamung von Wasser an hydrophoben Oberflächen bewiesen werden. Die gesammelten Ergebnisse und Erkenntnisse in diesen verschiedenen Themengebieten werden in Zukunft einen großen Einfluss in der Wissenschaftswelt haben. / In the presented dissertation new methods in the chain elongation of carbohydrates could be established and optimized. Besides that, new probes for the investigation of protonation and water dynamics could be designed and synthesized. The extension of comprehension in the interaction between amino acids and carbohydrates through hydrogen bonds has a great impact in the scientific community of carbohydrate research. The stereochemical control in the construction of stereotetrads and –pentads leads to a considerable extension of known methods in the synthesis of natural compounds and biomimetics. Additionally the C-glycosides of rare carbohydrates could be synthesized through simple methods. Installation of amino acids into carbohydrate structures could be achieved with very high stereoselectivity and the potential of manipulating the stereochemical course through the choice of different isocyanides. The synthesis of the cell permeable PAc-SNARF and the cysteine-bioapplicable IA-SNARF allow the ratiometric pH-imaging of cells and protein surfaces. The generalization of the synthesis of dye-precursors for SNARF-derivatives through friedel-crafts-acylation allow an inexpensive approach in synthesizing a broad spectrum of dyes. Through deployment of newly developed sterical demanding N-Methyl-6-oxyquinolinium betaine-derivates together with the fluorescence upconversion spectroscopy a deceleration of water reorientation near hydrophobic surfaces could be proven. The here summarized results and insights in the different topics will have a considerable influence in academic sciences.