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

Interactions between C-type lectin-like NK cell receptors and their protein ligands form one of the key recognition mechanisms of the innate immune system that is involved in the elimination of cells that have been malignantly transformed, virally infected, or stressed by chemotherapy or other factors. We determined an x-ray structure for the extracellular domain of mouse C-type lectin related (Clr) protein g, a ligand for the activation receptor NKR-P1F. Clr-g forms dimers in the crystal structure resembling those of human CD69. This newly reported structure, together with the previously determined structure of mouse receptor NKR-P1A, allowed the modeling and calculations of electrostatic profiles for other closely related receptors and ligands. Despite the high similarity among Clr-g, Clr-b, and human CD69, these molecules have fundamentally different electrostatics, with distinct polarization of Clr-g. The electrostatic profile of NKR-P1F is complementary to that of Clr-g, which suggests a plausible interaction mechanism based on contacts between surface sites of opposite potential.

• MeSH
• CD antigeny chemie   imunologie   MeSH
• diferenciační antigeny T-lymfocytů chemie   imunologie   MeSH
• krystalografie rentgenová MeSH
• lektiny typu C chemie   imunologie   MeSH
• lidé MeSH
• ligandy MeSH
• membránové proteiny chemie   imunologie   MeSH
• myši MeSH
• receptory imunologické chemie   imunologie   MeSH
• statická elektřina MeSH
• strukturní homologie proteinů MeSH
• terciární struktura proteinů MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• CD antigeny MeSH
• CD69 antigen MeSH Prohlížeč
• Dcl1 protein, mouse MeSH Prohlížeč
• diferenciační antigeny T-lymfocytů MeSH
• lektiny typu C MeSH
• ligandy MeSH
• membránové proteiny MeSH
• Nkrp1f protein, mouse MeSH Prohlížeč
• receptory imunologické MeSH