Oligomeric Architecture of Mouse Activating Nkrp1 Receptors on Living Cells

0505686 - MBÚ 2020 RIV CH eng J - Journal Article
Adámková, Ljubina - Kvíčalová, Zuzana - Rozbeský, Daniel - Kukačka, Zdeněk - Adámek, David - Cebecauer, Marek - Novák, Petr
Oligomeric Architecture of Mouse Activating Nkrp1 Receptors on Living Cells.
International Journal of Molecular Sciences. Roč. 20, č. 8 (2019), č. článku 1884. E-ISSN 1422-0067
R&D Projects: GA ČR(CZ) GA16-24309S; GA MŠMT(CZ) LH15010; GA MŠMT(CZ) LD15089; GA MŠMT(CZ) LQ1604; GA MŠMT(CZ) ED1.1.00/02.0109
Research Infrastructure: CIISB - 90043
Institutional support: RVO:61388971 ; RVO:61388955
Keywords : Nkrp1 * dimerization * Forster resonance energy transfer
OECD category: Biochemistry and molecular biology; Physical chemistry (UFCH-W)
Impact factor: 4.556, year: 2019
Method of publishing: Open access
https://www.mdpi.com/1422-0067/20/8/1884

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 Forster 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.
Permanent Link: http://hdl.handle.net/11104/0297120