Mouse activating Nkrp1 proteins are commonly described as type II transmembrane receptors with disulfide-linked homodimeric structure. Their function and the manner in which Nkrp1 proteins of mouse strain (C57BL/6) oligomerize are still poorly understood. To assess the oligomerization state of Nkrp1 proteins, mouse activating EGFP-Nkrp1s were expressed in mammalian lymphoid cells and their oligomerization evaluated by Förster resonance energy transfer (FRET). Alternatively, Nkrp1s oligomers were detected by Western blotting to specify the ratio between monomeric and dimeric forms. We also performed structural characterization of recombinant ectodomains of activating Nkrp1 receptors. Nkrp1 isoforms c1, c2 and f were expressed prevalently as homodimers, whereas the Nkrp1a displays larger proportion of monomers on the cell surface. Cysteine-to-serine mutants revealed the importance of all stalk cysteines for protein dimerization in living cells with a major influence of cysteine at position 74 in two Nkrp1 protein isoforms. Our results represent a new insight into the oligomerization of Nkrp1 receptors on lymphoid cells, which will help to determine their function.

• Klíčová slova
• Förster resonance energy transfer, Nkrp1, cysteine, dimerization, disulfide bond arrangement,
• MeSH
• antigeny Ly analýza   MeSH
• Cercopithecus aethiops MeSH
• COS buňky MeSH
• Jurkat buňky MeSH
• lektinové receptory NK-buněk - podrodina B analýza   chemie   MeSH
• lidé MeSH
• multimerizace proteinu MeSH
• myši inbrední C57BL MeSH
• myši MeSH
• receptory imunologické analýza   MeSH
• refolding proteinů MeSH
• rezonanční přenos fluorescenční energie MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• antigeny Ly MeSH
• Klrb1a protein, mouse MeSH Prohlížeč
• Klrb1c protein, mouse MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina B MeSH
• Nkrp1f protein, mouse MeSH Prohlížeč
• receptory imunologické MeSH

The C-type lectin-like receptors include the Nkrp1 protein family that regulates the activity of natural killer (NK) cells. Rat Nkrp1a was reported to bind monosaccharide moieties in a Ca2+-dependent manner in preference order of GalNac > GlcNAc >> Fuc >> Gal > Man. These findings established for rat Nkrp1a have been extrapolated to all additional Nkrp1 receptors and have been supported by numerous studies over the past two decades. However, since 1996 there has been controversy and another article showed lack of interactions with saccharides in 1999. Nevertheless, several high affinity saccharide ligands were synthesized in order to utilize their potential in antitumor therapy. Subsequently, protein ligands were introduced as specific binders for Nkrp1 proteins and three dimensional models of receptor/protein ligand interaction were derived from crystallographic data. Finally, for at least some members of the NK cell C-type lectin-like proteins, the "sweet story" was impaired by two reports in recent years. It has been shown that the rat Nkrp1a and CD69 do not bind saccharide ligands such as GlcNAc, GalNAc, chitotetraose and saccharide derivatives (GlcNAc-PAMAM) do not directly and specifically influence cytotoxic activity of NK cells as it was previously described.

• MeSH
• buňky NK * chemie   imunologie   metabolismus   MeSH
• CD antigeny * chemie   imunologie   metabolismus   MeSH
• diferenciační antigeny T-lymfocytů * chemie   imunologie   metabolismus   MeSH
• krysa rodu Rattus MeSH
• lektinové receptory NK-buněk - podrodina B * chemie   imunologie   metabolismus   MeSH
• lektiny typu C * chemie   imunologie   metabolismus   MeSH
• lidé MeSH
• oligosacharidy * chemie   imunologie   metabolismus   MeSH
• terciární struktura proteinů MeSH
• zvířata MeSH
• Check Tag
• krysa rodu Rattus MeSH
• lidé MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CD antigeny * MeSH
• CD69 antigen MeSH Prohlížeč
• diferenciační antigeny T-lymfocytů * MeSH
• KLRB1 protein, human MeSH Prohlížeč
• lektinové receptory NK-buněk - podrodina B * MeSH
• lektiny typu C * MeSH
• oligosacharidy * MeSH