A mucus-secreting human colonic cancer cell line. Purification and partial characterization of the secreted mucins
| Autor: | C Augeron, Jean-José Maoret, Christian L. Laboisse, Chantal Bauvy, Michèle Aubery, Patrice Codogno, J Font |
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| Rok vydání: | 1989 |
| Předmět: |
chemistry.chemical_classification
Gel electrophoresis biology Chemistry Mucin Mucins Cell Biology Adenocarcinoma Oligosaccharide Biochemistry Mucus Cell Line Immunoenzyme Techniques Cell culture Polyclonal antibodies Immunoblot Analysis Colonic Neoplasms Chromatography Gel Tumor Cells Cultured biology.protein Humans Glycoprotein Molecular Biology Research Article |
| Zdroj: | Biochemical Journal. 258:793-799 |
| ISSN: | 1470-8728 0264-6021 |
| DOI: | 10.1042/bj2580793 |
| Popis: | A stably differentiated clonal derivative (Cl.16E) of the human colonic adenocarcinoma cell line HT29 secretes in culture high-Mr glycoproteins that were purified from the serum-free conditioned medium by preparative SDS/polyacrylamide-gel electrophoresis. Analysis of the oligosaccharides released from the [3H]glucosamine-labelled high-Mr glycoproteins by alkaline-borohydride treatment showed that this material consisted of O-linked oligosaccharides (without any detectable N-linked oligosaccharides) that were eluted as three fractions from Bio-Gel P-6 columns. The main oligosaccharide fraction obtained after such treatment and desialylation was eluted together with a six-unit glucose polymer from a Bio-Gel P-4 column. Polyclonal antibodies were raised against the high-Mr glycoproteins, and in immunoblot analysis they reacted specifically with the high-Mr glycoproteins present in the conditioned medium. Furthermore, immunohistochemical staining of sections in paraffin wax revealed that these antibodies labelled normal human gastrointestinal mucins. We conclude that (1) the high-Mr glycoproteins prepared by SDS/polyacrylamide-gel electrophoresis are pure mucus glycoproteins on the basis of sensitivity to alkaline-borohydride treatment, monosaccharide composition and immunochemical and immunohistological findings, and (2) these mucins have antigenic determinants in common with the normal human gastrointestinal mucins. |
| Databáze: | OpenAIRE |
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