Počet záznamů: 1  

High-level expression and purification of soluble form of human natural killer cell receptor NKR-P1 in HEK293S GnTI(-) cells

  1. 1.
    SYSNO ASEP0489434
    Druh ASEPJ - Článek v odborném periodiku
    Zařazení RIVJ - Článek v odborném periodiku
    Poddruh JČlánek ve WOS
    NázevHigh-level expression and purification of soluble form of human natural killer cell receptor NKR-P1 in HEK293S GnTI(-) cells
    Tvůrce(i) Bláha, J. (CZ)
    Kalousková, B. (CZ)
    Skořepa, O. (CZ)
    Pažický, S. (CZ)
    Novák, Petr (MBU-M) RID, ORCID
    Vaněk, O. (CZ)
    Zdroj.dok.Protein Expression and Purification. - : Elsevier - ISSN 1046-5928
    Roč. 140, DEC 2017 (2017), s. 36-43
    Poč.str.8 s.
    Jazyk dok.eng - angličtina
    Země vyd.US - Spojené státy americké
    Klíč. slovaNKR-P1 ; CD161 ; klrb1
    Vědní obor RIVCE - Biochemie
    Obor OECDBiochemistry and molecular biology
    Institucionální podporaMBU-M - RVO:61388971
    UT WOS000411303200005
    EID SCOPUS85026659195
    DOI10.1016/j.pep.2017.07.016
    AnotaceHuman natural killer receptor protein 1 (NKR-P1, CD161, gene klrb1) is a C-type lectin-like receptor of natural killer (NK) cells responsible for recognition of its cognate protein ligand lectin-like transcript 1 (LLT1). NKR-P1 is the single human orthologue of the prototypical rodent NKR-P1 receptors. Naturally, human NKR-P1 is expressed on the surface of NK cells, where it serves as inhibitory receptor, and on T and NKT cells functioning as co-stimulatory receptor promoting secretion of IFN gamma. Most notably, it is expressed on Th17 and Tc17 lymphocytes where presumably promotes targeting into LLT1 expressing immunologically privileged niches. We tested effect of different protein tags (SUMO, TRX, GST, MsyB) on expression of soluble NKR-P1 in E. coli. Then we optimized the expression construct of soluble NKR-P1 by preparing a library of expression constructs in pOPING vector containing the extracellular lectin-like domain with different length of the putative N-terminal stalk region and tested its expression in Sf9 and HEK293 cells. Finally, a high-level expression of soluble NKR-P1 was achieved by stable expression in suspension-adapted HEK293S GnTI(-) cells utilizing pOPINGTTneo expression vector. Purified soluble NKR-P1 is homogeneous, deglycosylatable, crystallizable and monomeric in solution, as shown by size exclusion chromatography, multi-angle light scattering and analytical ultracentrifugation.
    PracovištěMikrobiologický ústav
    KontaktEliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
    Rok sběru2019
Počet záznamů: 1  

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