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Recombinant expression of rat NK cell receptor Clr-b in the presence of fluorinated analogues of monosaccharides
Urbanová, Zuzana ; Vaněk, Ondřej (advisor) ; Černá, Věra (referee)
NK cells are part of innate immunity that, besides eliminating damaged cells, also produce chemokines and cytokines, which affect the cells of adaptive immunity. NK cells express activating and inhibitory receptors on their surface. The balance between them keeps the NK cells inactive. When the balance is disrupted, the cytotoxic mechanisms of the cell are activated. Receptors NKR-P1B and NKR-P1D are two rat NK cell inhibitory receptors whose ligand is protein Clr-b, a receptor belonging to the C-type lectin-like receptor family. This work aimed to recombinantly produce Clr-b in the presence of seven fluorinated analogues of monosaccharides as potential inhibitors of N-glycosylation. The protein was successfully expressed in the HEK293T cell line as a construct containing the extracellular part of Clr-b, the Fc fragment of human IgG, and a histidine tag multiple times, each time in the presence of one of the compounds. As glycosylation plays a major role in the functionality of many proteins, inhibition of glycosylation appears to be a promising way of treatment of different diseases such as cancer or multiple sclerosis. The aim here was to assess the effect of fluorinated analogues of monosaccharides on the native N-glycosylation of the HEK293T cell line using the Clr-b construct as a model...
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