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

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F61388971%3A_____%2F17%3A00489434" target="_blank" >RIV/61388971:_____/17:00489434 - isvavai.cz</a>

Alternative codes found

RIV/00216208:11310/17:10360959

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>

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 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.

Czech name

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Czech description

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Type

J_imp - 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

Project

—

Continuities

I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2017

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

Protein Expression and Purification

ISSN

1046-5928

e-ISSN

—

Volume of the periodical

140

Issue of the periodical within the volume

DEC 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