New Alterations of Serum Glycoproteins in Alcoholic and Cirrhotic Patients Revealed by High Resolution Two-Dimensional Gel Electrophoresis

Gravel, Patricia; Walzer, Claude; Aubry, Christiane; Balant, L; Yersin, Bertrand; Hochstrasser, Denis F.; Guimón, José

doi:10.1006/bbrc.1996.0360

Cited by 70 publications

(40 citation statements)

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“…Genetic heterogeneity giving rise to reduced N-linked glycosylation of Bsep may be a contributing factor. A carbohydrate-deficient form of transferrin has been detected in alcoholic liver cirrhosis (6,14). Theoretically, decreased activity of N-linked glycosylation-related enzymes may give rise to insufficiently glycosylated Bsep, thereby impairing bile secretion.…”

Section: Discussionmentioning

confidence: 99%

TwoN-linked glycans are required to maintain the transport activity of the bile salt export pump (ABCB11) in MDCK II cells

Mochizuki¹,

Kagawa²,

Numari³

et al. 2007

American Journal of Physiology-Gastrointestinal and Liver Physi

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The aim of this study was to determine the role of N-linked glycosylation in protein stability, intracellular trafficking, and bile acid transport activity of the bile salt export pump [Bsep (ATP-binding cassette B11)]. Rat Bsep was fused with yellow fluorescent protein, and the following mutants, in which Asn residues of putative glycosylation sites (Asn109, Asn116, Asn122, and Asn125) were sequentially replaced with Gln, were constructed by site-directed mutagenesis: single N109Q, double N109Q + N116Q, triple N109Q + N116Q + N122Q, and quadruple N109Q + N116Q + N122Q + N125Q. Immunoblot and glycosidase cleavage analysis demonstrated that each site was glycosylated. Removal of glycans decreased taurocholate transport activity as determined in polarized MDCK II cells. This decrease resulted from rapid decay of the mutant Bsep protein; biochemical half-lives were 3.76, 3.65, 3.24, 1.35, and 0.52 h in wild-type, single-mutant, double-mutant, triple-mutant, and quadruple-mutant cells, respectively. Wild-type and single- and double-mutant proteins were distributed exclusively along the apical membranes, whereas triple- and quadruple-mutant proteins remained intracellular. MG-132 but not bafilomycin A1 extended the half-life, suggesting a role for the proteasome in Bsep degradation. To determine whether a specific glycosylation site or the number of glycans was critical for protein stability, we studied the protein expression of combinations of N-glycan-deficient mutants and observed that Bsep with one glycan was considerably unstable compared with Bsep harboring two or more glycans. In conclusion, at least two N-linked glycans are required for Bsep protein stability, intracellular trafficking, and function in the apical membrane.

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Section: Discussionmentioning

confidence: 99%

TwoN-linked glycans are required to maintain the transport activity of the bile salt export pump (ABCB11) in MDCK II cells

Mochizuki¹,

Kagawa²,

Numari³

et al. 2007

American Journal of Physiology-Gastrointestinal and Liver Physi

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“…Transferrin hypoglycosylation in these patients may be attributable to direct inhibition of galactosyltransferase activity by the accumulated galactose 1-phosphate or to an effect on the formation of UDP-galactose, the donorsubstrate in the reaction (27 ). Core 1 mucin type O-glycan (30,31 ). Plasma apoC-III was hypoglycosylated in the acute phase of S. pneumoniae-associated HUS in two patients.…”

Section: Discussionmentioning

confidence: 99%

Apolipoprotein C-III Isofocusing in the Diagnosis of Genetic Defects in O-Glycan Biosynthesis

Wopereis

Grünewald

Morava³

et al. 2003

Clinical Chemistry

131

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Background: Defects in the biosynthesis of N-glycans may be found by isoelectric focusing (IEF) of plasma transferrin. No test is available to demonstrate O-glycan biosynthesis defects. Methods: We used isoforms of apolipoprotein C-III (apoC-III) as a marker for the biosynthesis of core 1 mucin type O-glycans. Plasma samples from patients with primary defects and secondary alterations in Nglycan biosynthesis were studied by apoC-III isofocusing. Results: Age-related reference values for apoC-III were determined. Plasma samples from patients with the primary congenital disorders of glycosylation (CDG) types Ia-Ic, Ie, If, IIa, and IId all showed a normal apoC-III isofocusing profile. Plasma from two patients with CDG type IIx were tested: one showed a normal apoC-III distribution, whereas the other showed a hypoglycosylation profile. In plasma from patients with hemolytic uremic syndrome (HUS), a hypoglycosylation profile was obtained. Conclusions: IEF of apoC-III is a rapid and simple technique that may be used as a screening assay for

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“…Galectin-3-binding protein has about 7 N-glycosylation sites, which have substantial influence on antibody affinity; moreover, it is known that serum glycoproteins are 9 9 altered in alcoholic and cirrhotic patients [23,27,28]. In this regard, the role of N- (SuperSignal ® West Dura, Thermo Scientific) and imaging System (Versadoc, BioRad).…”

Section: Deglycosylation Of Serum Galectin-3-binding Proteinmentioning

confidence: 99%

“…Furthermore, we investigated the role of glycosylation in the difference of serum galectin-3-binding protein expression in 6 6 hepatitis C-and alcohol-related cirrhosis [17,23]. We chose cirrhosis for this comparison due to substantial difference in fibrogenesis and pathological scoring systems in pre-cirrhotic stages [24].…”

Section: Introductionmentioning

confidence: 99%

Galectin-3-binding protein: a serological and histological assessment in accordance with hepatitis C-related liver fibrosis

Cheung

Libbrecht²,

Tilleman³

et al. 2010

European Journal of Gastroenterology & Hepatology

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Objectives. Invasive liver biopsy is the current method for the assessment of liver fibrosis. In search of non-invasive alternatives, galectin-3-binding protein was introduced as a candidate-marker of hepatitis C-related fibrosis based on serum proteomics.We investigated the role of galectin-3-binding protein as a single-marker of significant fibrosis and cirrhosis by means of serology and histology and studied the role of glycosylation on the expression in hepatitis C-and alcohol-related cirrhosis. Methods. Sera and available biopsies of hepatitis C patients with various fibrosis-gradesand patients with alcohol-related cirrhosis were used for galectin-3-binding protein measurements by enzyme-linked immunosorbent assay and immunohistochemistry, respectively. Glycosylation-effect was analyzed by Western blot. Data was analyzed in accordance to fibrosis.Results. Galectin-3-binding protein was increased during cirrhosis (22.7 ± 10.1 µg/mL) in contrast to mild (11.3 ± 6.4 µg/mL) and moderate fibrosis (13.4 ± 8.3 µg/mL) (p < 0.001; p = 0.004, respectively). Receiver operator characteristics curves showed areas under the curve of 0.68 and 0.81 for the detection of respectively significant fibrosis and cirrhosis. Similar findings in hepatic expression were obtained, in which cirrhosis was associated with diffuse, parenchymal expression (p = 0.002). The observed difference between hepatitis C-and alcohol-related cirrhosis (13.5 ± 9.0 µg/mL) (p = 0.009) could not be explained by glycosylation. (mean ± standard deviation) Conclusions. Our recent findings confirm our initial proteome results on serological and histological level as well as the role of galectin-3-binding protein as a marker of 4 4 hepatitis C-related fibrosis, especially cirrhosis. Implication of this protein in future multi-marker study should be considered.

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New Alterations of Serum Glycoproteins in Alcoholic and Cirrhotic Patients Revealed by High Resolution Two-Dimensional Gel Electrophoresis

Cited by 70 publications

References 0 publications

TwoN-linked glycans are required to maintain the transport activity of the bile salt export pump (ABCB11) in MDCK II cells

TwoN-linked glycans are required to maintain the transport activity of the bile salt export pump (ABCB11) in MDCK II cells

Apolipoprotein C-III Isofocusing in the Diagnosis of Genetic Defects in O-Glycan Biosynthesis

Galectin-3-binding protein: a serological and histological assessment in accordance with hepatitis C-related liver fibrosis

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