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The C-type lectin-like receptor Nkrp1b: Structural proteomics reveals features affecting protein conformation and interactions

SYSNO ASEP	0508013
Druh ASEP	J - Článek v odborném periodiku
Zařazení RIV	J - Článek v odborném periodiku
Poddruh J	Článek ve WOS
Název	The C-type lectin-like receptor Nkrp1b: Structural proteomics reveals features affecting protein conformation and interactions
Tvůrce(i)	Hernychová, Lucie (MBU-M) Rosůlek, Michal (MBU-M)_ORCID Kádek, Alan (MBU-M)_{RID, ORCID} Mareška, V. (CZ) Chmelík, Josef (MBU-M ed.)_{RID, ORCID} Adámková, Ljubina (MBU-M) Grobárová, Valeria (MBU-M) Šebesta, O. (CZ) Kukačka, Zdeněk (MBU-M)_{RID, ORCID} Skála, Kristián (MBU-M) Spiwok, V. (CZ) Černý, J. (CZ) Novák, Petr (MBU-M)_{RID, ORCID}
Zdroj.dok.	Journal of Proteomics. - : Elsevier - ISSN 1874-3919 Roč. 196, MAR 30 (2019), s. 162-172
Poč.str.	11 s.
Jazyk dok.	eng - angličtina
Země vyd.	NL - Nizozemsko
Klíč. slova	Structural mass spectrometry ; Chemical cross-linking ; Ion mobility
Vědní obor RIV	CE - Biochemie
Obor OECD	Microbiology
CEP	GA16-24309S GA ČR - Grantová agentura ČR
	LQ1604 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy
	LM2015043 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy
	ED1.1.00/02.0109 GA MŠMT - Ministerstvo školství, mládeže a tělovýchovy
Výzkumná infrastruktura	Czech-BioImaging - 90062 - Ústav molekulární genetiky AV ČR, v. v. i.
Způsob publikování	Omezený přístup
Institucionální podpora	MBU-M - RVO:61388971
UT WOS	000460716800015
EID SCOPUS	85056670654
DOI	10.1016/j.jprot.2018.11.007
Anotace	The cytotoxicity of mouse natural killer (NK) cells in response to pathological changes in target cells is regulated via the Nkrp1b receptor. Here, we characterized the Nkrp1b structure and structural features (stalk, loop, and oligomerization state) that affect its interactions. To study the Nkrp1b protein structure and the functional importance of its stalk, two Nkrp1b protein variants differing by the presence of the stalk were prepared. These variants were studied using a combination of structural mass spectrometry approaches with computational modeling to derive structural models. In addition, information about biological activity and localization in mammalian cells was acquired using scanning microscopy techniques and western blotting. Based on these methods, we obtained the structure of Nkrp1b ectodomain in its monomeric and dimeric conformations, identified the dimerization interface, and determined disulfide connections within the molecule. We found that Nkrp1b occurs as a mixture of monomers and homodimers, both in vitro and in vivo. Significance: Despite the long-standing assumption that Nkrp1 proteins are homodimers connected by disulfide bonds in the stalk region, our data showed that both Nkrp1b protein variants form monomers and homodimers irrespective of the presence of the stalk. We demonstrated that the stalk is not crucial for protein dimerization or ligand binding and that Nkrp1b interacts with its natural ligands only in its monomeric conformation therefore, dimers may have another regulatory function. Using a unique combination of computational, biochemical, and biological methods, we revealed the structural conformation and behavior of Nkrp1b in its native state. In addition, it is a first report utilizing the intermolecular chemical cross-linking of light- and heavy-labeled protein chains together with ion mobility-mass spectrometry to design the structural models of protein homodimers.
Pracoviště	Mikrobiologický ústav
Kontakt	Eliška Spurná, eliska.spurna@biomed.cas.cz, Tel.: 241 062 231
Rok sběru	2020
Elektronická adresa	https://www.sciencedirect.com/science/article/pii/S187439191830397X?via%3Dihub

Počet záznamů: 1