High-level expression and purification of soluble form of human natural killer cell receptor NKR-P1 in HEK293S GnTI- cells
The result's identifiers
Result code in IS VaVaI
<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00216208%3A11310%2F17%3A10360959" target="_blank" >RIV/00216208:11310/17:10360959 - isvavai.cz</a>
Alternative codes found
RIV/61388971:_____/17:00489434
Result on the web
<a href="http://dx.doi.org/10.1016/j.pep.2017.07.016" target="_blank" >http://dx.doi.org/10.1016/j.pep.2017.07.016</a>
DOI - Digital Object Identifier
<a href="http://dx.doi.org/10.1016/j.pep.2017.07.016" target="_blank" >10.1016/j.pep.2017.07.016</a>
Alternative languages
Result language
angličtina
Original language name
High-level expression and purification of soluble form of human natural killer cell receptor NKR-P1 in HEK293S GnTI- cells
Original language description
Human 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 INFγ. 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 MINUS SIGN 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.
Czech name
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Czech description
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Classification
Type
J<sub>imp</sub> - Article in a specialist periodical, which is included in the Web of Science database
CEP classification
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OECD FORD branch
10608 - Biochemistry and molecular biology
Result continuities
Project
Result was created during the realization of more than one project. More information in the Projects tab.
Continuities
P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)
Others
Publication year
2017
Confidentiality
S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů
Data specific for result type
Name of the periodical
Protein Expression and Purification
ISSN
1046-5928
e-ISSN
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Volume of the periodical
140
Issue of the periodical within the volume
December 2017
Country of publishing house
US - UNITED STATES
Number of pages
8
Pages from-to
36-43
UT code for WoS article
000411303200005
EID of the result in the Scopus database
2-s2.0-85026659195