structural proteomics Dotaz Zobrazit nápovědu

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.

MeSH
lektinové receptory NK-buněk - podrodina B chemie metabolismus MeSH
molekulární modely * MeSH
multimerizace proteinu * MeSH
myši inbrední BALB C MeSH
myši MeSH
proteomika * MeSH
sekundární struktura proteinů MeSH
vztahy mezi strukturou a aktivitou MeSH
zvířata MeSH
Check Tag
myši MeSH
zvířata MeSH
Publikační typ
časopisecké články MeSH
práce podpořená grantem MeSH

Článek

Proteomics of survival structures of fungal pathogens

Fungal conidia and sclerotia could be considered the most important structures for pathogenesis as their ...

New biotechnology. 2016 ; 33 (5 Pt B) : 655-65. [pub] 20160108

N Biotechnol
ISSN 1876-4347
Medvik
Zdroj

Fungal pathogens are causal agents of numerous human, animal, and plant diseases. They employ various infection modes to overcome host defense systems. Infection mechanisms of different fungi have been subjected to many comprehensive studies. These investigations have been facilitated by the development of various '-omics' techniques, and proteomics has one of the leading roles in this regard. Fungal conidia and sclerotia could be considered the most important structures for pathogenesis as their germination is one of the first steps towards a host infection. They represent interesting objects for proteomic studies because of the presence of unique proteins with unexplored biotechnological potential required for pathogen viability, development and the subsequent host infection. Proteomic peculiarities of survival structures of different fungi, including those of biotechnological significance (e.g., Asperillus fumigatus, A. nidulans, Metarhizium anisopliae), in a dormant state, as well as changes in the protein production during early stages of fungal development are the subjects of the present review. We focused on biological aspects of proteomic studies of fungal survival structures rather than on an evaluation of proteomic approaches. For that reason, proteins that have been identified in this context are discussed from the point of view of their involvement in different biological processes and possible functions assigned to them. This is the first review paper summarizing recent advances in proteomics of fungal survival structures.

MeSH
biotechnologie MeSH
fungální proteiny fyziologie MeSH
houby růst a vývoj patogenita fyziologie MeSH
interakce hostitele a patogenu fyziologie MeSH
lidé MeSH
mycelium fyziologie MeSH
proteom fyziologie MeSH
proteomika metody MeSH
spory hub fyziologie MeSH
virulence fyziologie MeSH
zvířata MeSH
Check Tag
lidé MeSH
zvířata MeSH
Publikační typ
časopisecké články MeSH
přehledy MeSH

Kolekce

Publikováno

Filtry

structural proteomics Dotaz Zobrazit nápovědu

structural proteomics Dotaz Zobrazit nápovědu

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