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Statements

Subject Item
n2:RIV%2F00216208%3A11160%2F14%3A10293780%21RIV15-MSM-11160___
rdf:type
skos:Concept n10:Vysledek
rdfs:seeAlso
http://pubs.acs.org/doi/full/10.1021/pr4007798
dcterms:description
Although dendritic cells (DCs) control the priming of the adaptive immunity response, a comprehensive description of their behavior at the protein level is missing. The introduction of the into the field of DC research would therefore be highly beneficial. quantitative proteomic technique of metabolic labeling (SILAC) To achieve this, we applied SILAC labeling to primary bone marow-derived DCs (BMDCs). These cells combine both biological relevance and experimental feasibility, as their in vitro generation permits the use of C-13/N-15-labeled amino acids.. Interestingly, BMDCs appear to exhibit a very active arginine metabolism. Using standard cultivation conditions, similar to 20% of all protein-incorporated proline was a byproduct of heavy arginine degradation. In addition, the dissipation of N-15 from labeled arginine to the whole proteome was observed. The latter decreased the mass accuracy in MS and affected the natural isotopic distribution of peptides. SILAC-connected metabolic issues were shown to be enhanced by GM-CSF, which is used for the differentiation of DC progenitors. Modifications of the cultivation procedure suppressed the arginine-related effects, yielding cells with a proteome labeling efficiency of }= 90%. Importantly, BMDCs generated according to the new cultivation protocol preserved their resemblance to inflammatory DCs in vivo, as evidenced by their response to LPS treatment. Although dendritic cells (DCs) control the priming of the adaptive immunity response, a comprehensive description of their behavior at the protein level is missing. The introduction of the into the field of DC research would therefore be highly beneficial. quantitative proteomic technique of metabolic labeling (SILAC) To achieve this, we applied SILAC labeling to primary bone marow-derived DCs (BMDCs). These cells combine both biological relevance and experimental feasibility, as their in vitro generation permits the use of C-13/N-15-labeled amino acids.. Interestingly, BMDCs appear to exhibit a very active arginine metabolism. Using standard cultivation conditions, similar to 20% of all protein-incorporated proline was a byproduct of heavy arginine degradation. In addition, the dissipation of N-15 from labeled arginine to the whole proteome was observed. The latter decreased the mass accuracy in MS and affected the natural isotopic distribution of peptides. SILAC-connected metabolic issues were shown to be enhanced by GM-CSF, which is used for the differentiation of DC progenitors. Modifications of the cultivation procedure suppressed the arginine-related effects, yielding cells with a proteome labeling efficiency of }= 90%. Importantly, BMDCs generated according to the new cultivation protocol preserved their resemblance to inflammatory DCs in vivo, as evidenced by their response to LPS treatment.
dcterms:title
Application of SILAC Labeling to Primary Bone Marrow-Derived Dendritic Cells Reveals Extensive GM-CSF-Dependent Arginine Metabolism Application of SILAC Labeling to Primary Bone Marrow-Derived Dendritic Cells Reveals Extensive GM-CSF-Dependent Arginine Metabolism
skos:prefLabel
Application of SILAC Labeling to Primary Bone Marrow-Derived Dendritic Cells Reveals Extensive GM-CSF-Dependent Arginine Metabolism Application of SILAC Labeling to Primary Bone Marrow-Derived Dendritic Cells Reveals Extensive GM-CSF-Dependent Arginine Metabolism
skos:notation
RIV/00216208:11160/14:10293780!RIV15-MSM-11160___
n4:aktivita
n7:S n7:P n7:I
n4:aktivity
I, P(7AMB12GR038), S
n4:cisloPeriodika
2
n4:dodaniDat
n13:2015
n4:domaciTvurceVysledku
n11:1289020
n4:druhVysledku
n20:J
n4:duvernostUdaju
n19:S
n4:entitaPredkladatele
n17:predkladatel
n4:idSjednocenehoVysledku
3824
n4:idVysledku
RIV/00216208:11160/14:10293780
n4:jazykVysledku
n12:eng
n4:klicovaSlova
mass spectrometry; quantitative proteomics; lipopolysaccharide; peptide isotopic distribution; arginine metabolism; SILAC; dendritic cells
n4:klicoveSlovo
n8:quantitative%20proteomics n8:SILAC n8:peptide%20isotopic%20distribution n8:dendritic%20cells n8:lipopolysaccharide n8:mass%20spectrometry n8:arginine%20metabolism
n4:kodStatuVydavatele
US - Spojené státy americké
n4:kontrolniKodProRIV
[F94C70468B1B]
n4:nazevZdroje
Journal of Proteome Research
n4:obor
n18:CE
n4:pocetDomacichTvurcuVysledku
1
n4:pocetTvurcuVysledku
6
n4:projekt
n5:7AMB12GR038
n4:rokUplatneniVysledku
n13:2014
n4:svazekPeriodika
13
n4:tvurceVysledku
Fabrik, Ivo Härtlová, Anetta Stulík, Jiří Daňková, Věra Řehulka, Pavel Link, Marek
n4:wos
000331164100036
s:issn
1535-3893
s:numberOfPages
11
n14:doi
10.1021/pr4007798
n15:organizacniJednotka
11160