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Preparation of expression vectors for receptor NKp30 and its ligands B7-H6 and BAG-6
Pažický, Samuel ; Vaněk, Ondřej (advisor) ; Milichovský, Jan (referee)
NK cells, the cells of non-adaptive imune system, are able to recognise viraly infected or oncogenous cells by many inhibiting and activating receptors that are expressed on their surface and eliminate them consequently. For NKp30, activating receptor of NK cells included in NCR (Natural Killer Cell Receptors) family, lately there were identified several ligands, including membrane protein B7-H6 expressed on oncogenous cells surface and BAG- 6, cell core protein with wide spectrum of functions. Aim of this thesis was preparation of expression vectors coding for receptor NKp30 and his ligands B7-H6 and BAG-6, enabling expression of these proteins in HEK293 cell line. Keywords: NK cell, plasmid, receptor, NKp30, B7-H6, BAG-6
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Study of NKp30 oligomerization and its interaction with B7-H6
Pažický, Samuel ; Vaněk, Ondřej (advisor) ; Man, Petr (referee)
NK cells are important part of immune system, recognizing and eliminating tumor cells and cells infected by viruses. For the target cell recognition, binding of ligands by activating receptors plays a crucial role. Activating receptor NKp30, protein of family of natural cytotoxicity receptors, is able to bind multiple ligands either present on tumor cell surface or being part of some viruses. B7-H6 is one of the ligands of NKp30 and its specific constitutive expression on some tumor cells and cell lines makes it an interesting biological target. Although the NKp30/B7-H6 complex structure has been solved, structural basis of some important features of their binding is not explained yet. Soluble form of NKp30 receptor binding domain creates oligomers, presence of which is dependent on C-terminus length of its domain and its N-glycosylation; however, structural insight into formation of the oligomers and their significance is not known. Furthermore, binding affinity of NKp30 to its ligands is dependent on presence of its glycosylation and glycosylation type. We have already found out that NKp30 oligomerization is dependent on its glycosylation. In my work, I attempted to gain detailed functional and structural information about oligomerization of NKp30 and its binding to B7-H6 by multimethodical...
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Preparation of expression vectors for receptor NKp30 and its ligands B7-H6 and BAG-6
Pažický, Samuel ; Vaněk, Ondřej (advisor) ; Milichovský, Jan (referee)
NK cells, the cells of non-adaptive imune system, are able to recognise viraly infected or oncogenous cells by many inhibiting and activating receptors that are expressed on their surface and eliminate them consequently. For NKp30, activating receptor of NK cells included in NCR (Natural Killer Cell Receptors) family, lately there were identified several ligands, including membrane protein B7-H6 expressed on oncogenous cells surface and BAG- 6, cell core protein with wide spectrum of functions. Aim of this thesis was preparation of expression vectors coding for receptor NKp30 and his ligands B7-H6 and BAG-6, enabling expression of these proteins in HEK293 cell line. Keywords: NK cell, plasmid, receptor, NKp30, B7-H6, BAG-6
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