Enzymic Reconstruction of Glycosaminoglycan Oligosaccharide Chains Using the Transglycosylation Reaction of Bovine Testicular Hyaluronidase
| Autor: | Mitsuo Majima, Masahiko Endo, Hozumi Narita, Hiromi Saitoh, Akihiko Matsuki, Masaharu Kasai, Keiichi Takagaki, Toshiya Nakamura |
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| Rok vydání: | 1995 |
| Předmět: |
Male
Glycosylation Stereochemistry Molecular Sequence Data Disaccharide Hyaluronoglucosaminidase Oligosaccharides Sodium Chloride Biochemistry Glycosaminoglycan chemistry.chemical_compound Hydrolysis Testis Carbohydrate Conformation Animals Chondroitin Molecular Biology Glycosaminoglycans chemistry.chemical_classification Cell Biology Hydrogen-Ion Concentration Oligosaccharide Glucuronic acid Kinetics Carbohydrate Sequence chemistry Cattle Carbohydrate conformation |
| Zdroj: | Journal of Biological Chemistry. 270:3741-3747 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.270.8.3741 |
| Popis: | The reconstruction of glycosaminoglycan chains using the transglycosylation reaction of testicular hyaluronidase was investigated. First, the optimal conditions for the transglycosylation reaction catalyzed by the enzyme were determined by incubation with the enzyme, using hyaluronic acid (M(r) = 800,000) as a donor and pyridylaminated hyaluronic acid hexasaccharide having glucuronic acid at the nonreducing terminal as an acceptor. The carbohydrate chains as reaction products were determined by high performance liquid chromatography and mass spectrometry. The optimal pH for hydrolysis by the enzyme was found to be about 5.0, whereas that for the transglycosylation reaction was about 7.0. Sodium chloride in the reaction medium inhibited the transglycosylation reaction. Under the optimal conditions, the carbohydrate chains were sequentially transferred along with disaccharide units to the nonreducing terminal of the acceptor and elongated up to docosasaccharide from the acceptor, pyridylaminated hexasaccharide. Using a combination of hyaluronic acid, chondroitin, and chondroitin 4- and 6-sulfate as an acceptor and a donor, it was possible to reconstruct hybrid chains, which were natural or unnatural types of glycosaminoglycan chains. Therefore, it is highly likely that application of the transglycosylation reaction using testicular hyaluronidase would facilitate artificial reconstruction of glycosaminoglycans having some physiological functions. |
| Databáze: | OpenAIRE |
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