Selective release of the disaccharide 2-acetamido-2-deoxy-3-O-(β-d-galactopyranosyl)-d-galactose from epiglycanin by endo-N-acetyl-α-d-galactosaminidase
| Autor: | Veerasingham P. Bhavanandan, John F. Codington |
|---|---|
| Rok vydání: | 1983 |
| Předmět: |
Disaccharide
Disaccharides medicine.disease_cause Biochemistry alpha-N-Acetylgalactosaminidase Analytical Chemistry chemistry.chemical_compound medicine Animals Antigens Tumor-Associated Carbohydrate Amino Acids Glycoproteins chemistry.chemical_classification Membrane Glycoproteins Chromatography Organic Chemistry Galactitol Mammary Neoplasms Experimental Hexosamines General Medicine Clostridium perfringens Oligosaccharide Hexosaminidases Enzyme chemistry alpha-Galactosidase Galactose Tritium Glycoprotein |
| Zdroj: | Carbohydrate Research. 118:81-89 |
| ISSN: | 0008-6215 |
| DOI: | 10.1016/0008-6215(83)88037-9 |
| Popis: | Epiglycanin, the major glycoprotein of TA3-Ha mammary carcinoma ascites cells, was radiolabeled with tritium in the terminal d -galactose and 2-acetamido-2-deoxy- d -galactose residues. Alkaline-borohydride treatment, reported to release five O-glycosyl-linked chain types from epiglycanin, resulted in the cleavage of 98–99% of the radioactivity from the protein. Of this, 63% of the radioactivity from epiglycanin and 70% from asialoepiglycanin co-migrated with an authentic sample of 2-acetamido-2-deoxy-3-O-(β- d -galactopyranosyl)- d -galactitol on a column of Bio-Gel P-6. Incubation of [3H]galactose-epiglycanin with endo-N-acetyl -α- d -galactosaminidase (Diplococcus pneumoniae), and fractionation of the mixture on a column of Bio-Gel P-4, gave only one oligosaccharide peak containing 62 and 70%, respectively, of the radioactivity of epiglycanin and asialoepiglycanin. This oligosaccharide comigrated with authentic 2-acetamido-2-deoxy-3-O-(β- d -galactopyranosyl)- d -galactose (1) on columns of Bio-Gel P-2 and P-4 and on paper chromatograms. Results of experiments in which unlabeled epiglycanin was treated with enzyme and the products analyzed, by three different methods, suggested that 78–85% of 1 had been cleaved. Another enzyme, N-acetyl-α- d -galactosaminyl-oligosaccharidase from Clostridium perfringens, exhibited similar specificity and cleaved 65% of the radioactivity from ([3H]galactose)asialoepiglycanin, which was eluted from a Bio-Gel P-2 column as the disaccharide 1. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|