Structural differences between complex-type Asn-linked glycan chains of glycoproteins in rat hepatocytes and Zajdela hepatoma cells

Goulut-Chassaing, Chantal; Bourrillon, R

doi:10.1016/0304-4165(94)00191-y

Cited by 11 publications

(7 citation statements)

References 38 publications

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“…In rat hepatocytes oversialylation of biantennary N-glycan has been described only on the α1,3-bound NeuAcα2,3Galβ1,3Glc-NAcβ1,2Man-antennae of the biantennary N-glycan (Goulut-Chassaing and Bourrillon, 1995). Spik et al (1991) and Baussant et al (1992) also detected oversialylation in biantennary N-glycans of rat plasma glycoproteins in type I chains attached to the α1,3-linked mannose.…”

Section: Discussionmentioning

confidence: 98%

N-Glycosylation of the carcinoembryonic antigen related cell adhesion molecule, C-CAM, from rat liver: detection of oversialylated bi- and triantennary structures

et al. 1999

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Rat C-CAM is a ubiquitous, transmembrane and carcinoembryonic antigen related cell adhesion molecule. The human counterpart is known as biliary glycoprotein (BGP) or CD66a. It is involved in different cellular functions ranging from intercellular adhesion, microbial receptor activity, signaling and tumor suppression. In the present study N-glycosylation of C-CAM immunopurified from rat liver was analyzed in detail. The primary sequence of rat C-CAM contains 15 potential N-glycosylation sites. The N-glycans were enzymatically released from glycopeptides, fluorescently labeled with 2-aminobenzamide, and separated by two-dimensional HPLC. Oligosaccharide structures were characterized by enzymatic sequencing and MALDI-TOF-MS. Mainly bi- and triantennary complex structures were identified. The presence of type I and type II chains in the antennae of these glycans results in heterogeneous glycosylation of C-CAM. Sialylation of the sugars was found to be unusual; bi- and triantennary glycans contained three and four sialic acid residues, respectively, and this linkage seemed to be restricted to the type I chain in the antennae. Approximately 20% of the detected sugars contain these unusual numbers of sialic acids. C-CAM is the first transmembrane protein found to be oversialylated.

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Section: Discussionmentioning

confidence: 98%

N-Glycosylation of the carcinoembryonic antigen related cell adhesion molecule, C-CAM, from rat liver: detection of oversialylated bi- and triantennary structures

et al. 1999

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“…In choriocarcinoma cell lines, a cancer of the uterus, H 5 N 5 F 1 A 2 , containing a bisecting GlcNac, was uniquely observed compared to normal cell lines. (61) H 5 N 5 F 1 A 2 was expressed abnormally in hepatocytes,(62) which could be indicative of liver secretions triggered in response to OVC. Interestingly, we could not find any studies in cancer that identified the H 8 N 7 A 2 glycan, making it a potentially novel candidate.…”

Section: Discussionmentioning

confidence: 99%

Relative Quantification and Higher-Order Modeling of the Plasma Glycan Cancer Burden Ratio in Ovarian Cancer Case-Control Samples

Hecht¹,

Scholl²,

Walker³

et al. 2015

J. Proteome Res.

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An early-stage, population-wide biomarker for ovarian cancer (OVC) is essential to reverse its high mortality rate. Aberrant glycosylation by OVC has been reported, but studies have yet to identify an N-glycan with sufficiently high specificity. We curated a human biorepository of 82 case-control plasma samples, with 27%, 12%, 46%, and 15% falling across stages I–IV, respectively. For relatve quantitation, glycans were analyzed by the individuality normalization when labeling with glycan hydrazide tags (INLIGHT) strategy for enhanced electrospray ionization, MS/MS analysis. Sixty-three glycan cancer burden ratios (GBRs), defined as the log10 ratio of the case-control extracted ion chromatogram abundances, were calculated above the limit of detection. The final GBR models, built using stepwise forward regression, included three significant terms: OVC stage, normalized mean GBR, and tag chemical purity; glycan class, fucosylation, or sialylation were not significant variables. After Bonferroni correction, seven N-glycans were identified as significant (p < 0.05), and after false discovery rate correction, an additional four glycans were determined to be significant (p < 0.05), with one borderline (p = 0.05). For all N-glycans, the vectors of the effects from stages II–IV were sequentially reversed, suggesting potential biological changes in OVC morphology or in host response.

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“…Delipidated cell extracts or membrane glycoprotein fractions were treated with pronase (Calbiochem) as described previously [31]. Pronase digests were dissolved in 1 ml 100 mM pyridine-acetate, pH 5.3, and glycopeptides were separated by chromatography on a Bio-Gel P6 column (Bio-Rad).…”

Section: Glycoprotein Isolation By Electroelution Bands Identifiedmentioning

confidence: 99%

“…Here, tumor cells were maintained in the ascites form by serial intraperitoneal transplantation of the cells in Sprague Dawley rats 1271. Normal hepatocytes were prepared from Sprague Dawley rats by collagenase perfusion of the liver Zajdela hepatoma cells and normal hepatocytes were cultured as described previously [31], in the presence (1 pg/ml) or in the absence of tunicamycin (Glaxo Group Research Ltd), an inhibitor of the N-glycosylation process. After a 4-h incubation, cells were labeled for 24 h in the appropriate media with either ~-[2-'H]mannose (50 kBq/106 cells) or ~-[6-'H]glucosamine (I00 kBq/106 cells) (NEN).…”

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Expression and Characterization of a Lactosaminoglycan‐Carrying Glycoprotein of Zajdela Hepatoma Cell Surface

Goulut-Chassaing

Bourrillon

1997

European Journal of Biochemistry

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In poorly differentiated hepatoma cells, a glycoprotein carrying lactosaminoglycans is identified, and the structure of its glycan moiety is proposed. After membrane solubilization, protein fractionation by gel filtration, and electroelution, this glycoprotein (GPIII) was identified by its affinity for Datura stramonium lectin and its content in large glycopeptides. As shown by PAGE, GPIII has an apparent molecular mass of 100 kDa and is highly glycosylated (36%). It appears as an integral membrane glycoprotein. It is absent from normal hepatocytes, in that no heavy glycopeptides could be detected that bound to Datura lectin or to specific antiserum. The glycan moiety of GPIII has been analyzed according to carbohydrate composition, glycosidase treatment, affinity chromatography on immobilized pokeweed, Datura and Griffonia lectins, and by NMR and methylation analyses. The glycan is a N-linked tetraantennary lactosaminoglycan of 6.6 kDa, containing Gal, GlcNAc, Man, and NeuNAc in a 16: 14:3:4 molar ratio, with an average of three repeating unitshranch. Its P-Gal residues are in the penultimate position and are linked in PI-4 at least in four structural elements (three peripheral and one internal). It contains a very branched structure with Galal-3Gal/31-4GlcNAc side chains linked in the C6 position to an inner Gal residue in a main branch. a-Gal and NeuNAc residues [mainly NeuNAca(2-3) linkage] are expressed as the nonreducing terminal groups. A possible structural model is proposed for this heterogeneous lactosaminoglycan, although no definitive structure can be established. That this lactosaminoglycan-carrying glycoprotein GPIII is not expressed in hepatocytes suggests its expression to be linked to the undifferentiated and/or malignant state of this hepatoma.Keywords: membrane glycoprotein; protein carrier; malignant transformation; hepatoma cells ; N-glycan chain.Lactosaminoglycans are high-molecular-mass glycans that consist of Gal(~1-4GlcNAc@1-3) repeating units, attached as linear or branched structures to a (Man),(GlcNAc), core. They are sensitive to endo P-galactosidase, and differ either in their number of repeating units or through the presence of branching points in GalV1-6) or in the nature of their non-reducing terminal group (either NeuAc, a-Gal or P-Gal) [l -41.Initially discovered and characterized in erythrocytes [I -3, 5, 61, lactosaminoglycans are also expressed in significant amounts in the hematopoietic systems [7-91, and at the surface of embryonic [lO-12] and transformed cell lines [13-181, and are described in limited amounts in normal adult tissues [2, 171. Furthermore these structures display dramatic changes in their branching patterns and peripheral substitutions during cell differentiation, oncogenesis and embryonic development. Although a relationship seems to exist between these alterations in lactosaminoglycan patterns and the biological processes [4,12, 191, the function of these glycans remains unknown.Correspondence to R. Bourrillon, U.F.R. BiomCdicale des Saints- Although di...

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Structural differences between complex-type Asn-linked glycan chains of glycoproteins in rat hepatocytes and Zajdela hepatoma cells

Cited by 11 publications

References 38 publications

N-Glycosylation of the carcinoembryonic antigen related cell adhesion molecule, C-CAM, from rat liver: detection of oversialylated bi- and triantennary structures

N-Glycosylation of the carcinoembryonic antigen related cell adhesion molecule, C-CAM, from rat liver: detection of oversialylated bi- and triantennary structures

Relative Quantification and Higher-Order Modeling of the Plasma Glycan Cancer Burden Ratio in Ovarian Cancer Case-Control Samples

Expression and Characterization of a Lactosaminoglycan‐Carrying Glycoprotein of Zajdela Hepatoma Cell Surface

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