0440676 - MBÚ 2015 RIV US eng J - Journal Article
Pompach, Petr - Ashline, David J. - Brnáková, Z. - Benicky, J. - Sanda, M. - Goldman, R.
Protein and Site Specificity of Fucosylation in Liver-Secreted Glycoproteins.
Journal of Proteome Research. Roč. 13, č. 12 (2014), s. 5561-5569. ISSN 1535-3893. E-ISSN 1535-3907
R&D Projects: GA MŠMT LH13051; GA ČR GAP206/12/0503
Grant - others:Charles Univ.(CZ) UNCE_204025/2012
Institutional support: RVO:61388971
Keywords : fucose * glycoproteins * liver * site specificity
Subject RIV: CE - Biochemistry
Impact factor: 4.245, year: 2014

Chronic liver diseases are a serious health problem worldwide. One of the frequently reported glycan alterations in liver disease is aberrant fucosylation, which was suggested as a marker for noninvasive serologic monitoring. We present a case study that compares site specific glycoforms of four proteins including haptoglobin, complement factor H, kininogen-1, and hemopexin isolated from the same patient. Our exoglycosidase-assisted LC-MS/MS analysis confirms the high degree of fucosylation of some of the proteins but shows that microheterogeneity is protein- and site-specific. MSn analysis of permethylated detached glycans confirms the presence of LeY glycoforms on haptoglobin, which cannot be detected in hemopexin or complement factor H; all three proteins carry Lewis and H epitopes. Core fucosylation is detectable in only trace amounts in haptoglobin but with confidence on hemopexin and complement factor H, where core fucosylation of the bi-antennary glycans on select glycopeptides reaches 15-20% intensity. These protein-specific differences in fucosylation, observed in proteins isolated from the same patient source, suggest that factors other than up-regulation of enzymatic activity regulate the microheterogeneity of glycoforms. This has implications for selection of candidate proteins for disease monitoring and suggests that site-specific glycoforms have structural determinants, which could lead to functional consequences for specific subsets of proteins or their domains.
Permanent Link: http://hdl.handle.net/11104/0243777