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Statements

Subject Item
n2:RIV%2F61388971%3A_____%2F14%3A00440635%21RIV15-GA0-61388971
rdf:type
skos:Concept n16:Vysledek
dcterms:description
Enhanced fucosylation has been suggested as a marker for serologic monitoring of liver disease and hepatocellular carcinoma (HCC). We present a workflow for quantitative site-specific analysis of fucosylation and apply it to a comparison of hemopexin (HPX) and complement factor H (CFH), two liver-secreted glycoproteins, in healthy individuals and patients with liver cirrhosis and HCC. Label-free LC-MS quantification of glycopeptides derived from these purified glycoproteins was performed on pooled samples (2 pools/group, 5 samples/pool) and complemented by glycosidase assisted analysis using sialidase and endoglycosidase F2/F3, respectively, to improve resolution of glycoforms. Our analysis, presented as relative abundance of individual fucosylated glycoforms normalized to the level of their nonfucosylated counterparts, revealed a consistent increase in fucosylation in liver disease with significant site- and protein-specific differences. We have observed the highest microheterogeneity of glycoforms at the N187 site of HPX, absence of core fucosylation at N882 and N911 sites of CFH, or a higher degree of core fucosylation in CFH compared to HPX, but we did not identify changes differentiating HCC from matched cirrhosis samples. Glycosidase assisted LC-MS-MRM analysis of individual patient samples prepared by a simplified protocol confirmed the quantitative differences. Transitions specific to outer arm fucose document a disease-associated increase in outer arm fucose on both bi- and triantennary glycans at the N187 site of HPX. Further verification is needed to confirm that enhanced fucosylation of HPX and CFH may serve as an indicator of premalignant liver disease. The analytical strategy can be readily adapted to analysis of other proteins in the appropriate disease context. Enhanced fucosylation has been suggested as a marker for serologic monitoring of liver disease and hepatocellular carcinoma (HCC). We present a workflow for quantitative site-specific analysis of fucosylation and apply it to a comparison of hemopexin (HPX) and complement factor H (CFH), two liver-secreted glycoproteins, in healthy individuals and patients with liver cirrhosis and HCC. Label-free LC-MS quantification of glycopeptides derived from these purified glycoproteins was performed on pooled samples (2 pools/group, 5 samples/pool) and complemented by glycosidase assisted analysis using sialidase and endoglycosidase F2/F3, respectively, to improve resolution of glycoforms. Our analysis, presented as relative abundance of individual fucosylated glycoforms normalized to the level of their nonfucosylated counterparts, revealed a consistent increase in fucosylation in liver disease with significant site- and protein-specific differences. We have observed the highest microheterogeneity of glycoforms at the N187 site of HPX, absence of core fucosylation at N882 and N911 sites of CFH, or a higher degree of core fucosylation in CFH compared to HPX, but we did not identify changes differentiating HCC from matched cirrhosis samples. Glycosidase assisted LC-MS-MRM analysis of individual patient samples prepared by a simplified protocol confirmed the quantitative differences. Transitions specific to outer arm fucose document a disease-associated increase in outer arm fucose on both bi- and triantennary glycans at the N187 site of HPX. Further verification is needed to confirm that enhanced fucosylation of HPX and CFH may serve as an indicator of premalignant liver disease. The analytical strategy can be readily adapted to analysis of other proteins in the appropriate disease context.
dcterms:title
Quantification of Fucosylated Hemopexin and Complement Factor H in Plasma of Patients with Liver Disease Quantification of Fucosylated Hemopexin and Complement Factor H in Plasma of Patients with Liver Disease
skos:prefLabel
Quantification of Fucosylated Hemopexin and Complement Factor H in Plasma of Patients with Liver Disease Quantification of Fucosylated Hemopexin and Complement Factor H in Plasma of Patients with Liver Disease
skos:notation
RIV/61388971:_____/14:00440635!RIV15-GA0-61388971
n3:aktivita
n17:P n17:I
n3:aktivity
I, P(GAP206/12/0503), P(LH13051)
n3:cisloPeriodika
21
n3:dodaniDat
n5:2015
n3:domaciTvurceVysledku
n4:4576462
n3:druhVysledku
n15:J
n3:duvernostUdaju
n13:S
n3:entitaPredkladatele
n12:predkladatel
n3:idSjednocenehoVysledku
41058
n3:idVysledku
RIV/61388971:_____/14:00440635
n3:jazykVysledku
n7:eng
n3:klicovaSlova
fucose; hemopexin; complement
n3:klicoveSlovo
n10:fucose n10:complement n10:hemopexin
n3:kodStatuVydavatele
NL - Nizozemsko
n3:kontrolniKodProRIV
[A54F68706F20]
n3:nazevZdroje
Analytical Chemistry
n3:obor
n11:CB
n3:pocetDomacichTvurcuVysledku
1
n3:pocetTvurcuVysledku
5
n3:projekt
n8:LH13051 n8:GAP206%2F12%2F0503
n3:rokUplatneniVysledku
n5:2014
n3:svazekPeriodika
86
n3:tvurceVysledku
Wu, J. Sanda, M. Pompach, Petr Goldman, R. Benicky, J.
n3:wos
000344510200032
s:issn
0793-0135
s:numberOfPages
8