Recombinant expression of the mouse NK cell receptor NKR-P1C

NK cells are at the forefront of the body's defense against various threats, such as viruses or cancer cells. NK cells can recognize and destroy harmful cells in several ways that are very different from the defense mechanisms of, for example, T- or B-lymphocytes. NK cells carry a wide range of activating and inhibitory receptors on their surface that distinguish healthy cells from harmful or infected cells. NKR-P1 (natural killer receptor-protein 1) is the first discovered family of NK cell receptors similar to C-type lectins, which includes both activating and inhibitory receptors. NKR-P1C is an activating receptor capable of forming homodimers on the surface of NK cells. The major ligands of the NKR-P1 receptor family are the immunoenvasins of murine cytomegalovirus and other molecules structurally distinct from MHC class I glycoproteins. This work focuses on the recombinant expression of the NKR-P1C receptor protein. Using the pUT7 plasmid, we prepared a stably transfected HEK293T cell line carrying the gene for expression of the extracellular domain of NKR-P1C fused to the Fc fragment of IgG using the PiggiyBac system, and between them, they carry the TEV protease cleavage site. The protein was purified by affinity and gel permeation chromatography, and its purity was subsequently verified by..