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Synthesis of "Chemical Tags" and their application for selective protein labelling
Prokešová, Kristína ; Miletín, Miroslav (advisor) ; Nováková, Veronika (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Student: Kristína Prokešová Supervisor: doc. PharmDr. Miroslav Miletín, PhD. Consultant: Dr. Richard Wombacher Title: Synthesis of "Chemical Tags" and their application for selective protein labelling This diploma thesis is aimed on the synthesis and application of chemical tags. In theoretical part, protein labelling in general is discussed and fluorescent proteins, as routinely used technique for protein tracking, are shortly presented. The greatest attention is dedicated to chemical tags which consist of genetically encoded protein or peptide tag fused to the protein of interest (POI) and from a small fluorescent molecule which labels the POI-tag fusion. Particular representatives with their advantageous and disadvantageous properties are mentioned and super-resolution microscopy and calcium imaging, as applications of chemical tags, are explained. Experimental part is divided into chemical synthesis and biological methods. In synthetic part, four precursors of chemical tags have been prepared - two precursors of TMP-tag small molecule and two precursors of a chemical calcium dye. These precursors can be connected to create a new TMP-tag applicable for calcium imaging. One...
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Synthesis of "Chemical Tags" and their application for selective protein labelling
Prokešová, Kristína ; Miletín, Miroslav (advisor) ; Nováková, Veronika (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Pharmaceutical Chemistry and Drug Control Student: Kristína Prokešová Supervisor: doc. PharmDr. Miroslav Miletín, PhD. Consultant: Dr. Richard Wombacher Title: Synthesis of "Chemical Tags" and their application for selective protein labelling This diploma thesis is aimed on the synthesis and application of chemical tags. In theoretical part, protein labelling in general is discussed and fluorescent proteins, as routinely used technique for protein tracking, are shortly presented. The greatest attention is dedicated to chemical tags which consist of genetically encoded protein or peptide tag fused to the protein of interest (POI) and from a small fluorescent molecule which labels the POI-tag fusion. Particular representatives with their advantageous and disadvantageous properties are mentioned and super-resolution microscopy and calcium imaging, as applications of chemical tags, are explained. Experimental part is divided into chemical synthesis and biological methods. In synthetic part, four precursors of chemical tags have been prepared - two precursors of TMP-tag small molecule and two precursors of a chemical calcium dye. These precursors can be connected to create a new TMP-tag applicable for calcium imaging. One...
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Bifunctional BODIPY for optical labelling of biomolecules
Bartoň, Jan ; Kotek, Jan (advisor) ; Henke, Petr (referee)
The water-soluble derivate of BODIPY was prepared, which will be further modified in order to prepare bioorthogonal bifunctional BODIPY. Target application of this derivate is fluorescent probe for labelling of biomolecules. Main goals of this thesis were optimalization of synthesis of BODIPY core and sulfonation to positions 2,6. It was found out that sodium salt of sulfonated BODIPY shows good solublility in water and methanol, but it is poorly soluble in less polar solvents. Based on these findings DIPEA salt of sulfonated BODIPY was prepared, which shows outstanding range of solubility from water to dichlormethane. These findings will be used for synthesis of bifunctional water- soluble BODIPY. Further, the potential of Pd-BODIPY complex for detection of CO was reasserted which will be further investigated. Keywords BODIPY, bifunctional, water-soluble, fluorescent probe, fluorescence, protein labelling, solubilization
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