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Production of recombinant soluble dimeric C-type lectin-like receptors of rat natural killer cells
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SYSNO ASEP 0521615 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Production of recombinant soluble dimeric C-type lectin-like receptors of rat natural killer cells Author(s) Vaněk, O. (CZ)
Celadová, P. (CZ)
Skořepa, O. (CZ)
Bláha, J. (CZ)
Kalousková, B. (CZ)
Dvorská, A. (CZ)
Poalchová, E. (CZ)
Pucholtová, H. (CZ)
Kavan, Daniel (MBU-M) RID, ORCID
Pompach, Petr (MBU-M) RID, ORCID
Hofbauerová, Kateřina (MBU-M) ORCID
Kopecký, V. (CZ)
Mesci, A. (CA)
Voigt, S. (DE)
Carlyle, J. (CA)Article number 17836 Source Title Scientific Reports. - : Nature Publishing Group - ISSN 2045-2322
Roč. 9, NOV28 (2019)Number of pages 16 s. Language eng - English Country GB - United Kingdom Keywords missing-self-recognition ; secondary structure ; infrared-spectroscopy Subject RIV EE - Microbiology, Virology OECD category Microbiology Method of publishing Open access Institutional support MBU-M - RVO:61388971 UT WOS 000499236800001 EID SCOPUS 85075674332 DOI 10.1038/s41598-019-52114-8 Annotation Working at the border between innate and adaptive immunity, natural killer (NK) cells play a key role in the immune system by protecting healthy cells and by eliminating malignantly transformed, stressed or virally infected cells. NK cell recognition of a target cell is mediated by a receptor 'zipper' consisting of various activating and inhibitory receptors, including C-type lectin-like receptors. Among this major group of receptors, two of the largest rodent receptor families are the NKR-P1 and the Clr receptor families. Although these families have been shown to encode receptor-ligand pairs involved in MHC-independent self-nonself discrimination and are a target for immune evasion by tumour cells and viruses, structural mechanisms of their mutual recognition remain less well characterized. Therefore, we developed a non-viral eukaryotic expression system based on transient transfection of suspension-adapted human embryonic kidney 293 cells to produce soluble native disulphide dimers of NK cell C-type lectin-like receptor ectodomains. The expression system was optimized using green fluorescent protein and secreted alkaline phosphatase, easily quantifiable markers of recombinant protein production. We describe an application of this approach to the recombinant protein production and characterization of native rat NKR-P1B and Clr-11 proteins suitable for further structural and functional studies. Workplace Institute of Microbiology Contact Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231 Year of Publishing 2020 Electronic address https://www.nature.com/articles/s41598-019-52114-8
Number of the records: 1