A new type of exo-beta-glucuronidase acting only on non-sulfated glycosaminoglycans
| Autor: | Keiichi Takagaki, Kohmei Kubo, S Kimura, Mitsuo Majima, Toshiya Nakamura, M Endoss |
|---|---|
| Rok vydání: | 1990 |
| Předmět: |
Male
Immunodiffusion Chemical Phenomena Size-exclusion chromatography Oligosaccharides Sodium Chloride Biochemistry Substrate Specificity chemistry.chemical_compound Sulfation Affinity chromatography Animals Chondroitin Molecular Biology Polyacrylamide gel electrophoresis Glucuronidase Glycosaminoglycans chemistry.chemical_classification Gel electrophoresis Chromatography Chemistry Physical Sulfates Chemistry Substrate (chemistry) Rats Inbred Strains Cell Biology Rats Molecular Weight Enzyme Liver Ammonium Sulfate Electrophoresis Polyacrylamide Gel Rabbits |
| Zdroj: | Journal of Biological Chemistry. 265:5390-5397 |
| ISSN: | 0021-9258 |
| DOI: | 10.1016/s0021-9258(19)39371-8 |
| Popis: | Using chondroitin as a substrate, a new type of exo-beta-glucuronidase (EC 3.2.1.31) from rabbit liver was purified using a combination of ammonium sulfate fractionation, DEAE-cellulose chromatography, gel filtration on Sephracryl S-300, affinity chromatography through heparin-Sepharose CL-6B, and preparative polyacrylamide gel electrophoresis. This enzyme acts only on non-sulfated glycosaminoglycans and their oligosaccharides and was shown to be quite different from exo-beta-glucuronidase, which does act on p-nitro-phenyl-beta-D-glucuronide with regard to the following properties. 1) Neither sulfated glycosaminoglycanoligosaccharides nor p-nitrophenyl-beta-D-glucuronide were substrates for the enzyme. 2) The molecular weight was found to be about 130,000 by gel filtration, compared with a molecular weight of 280,000-300,000 for beta-glucuronidase, which acts on p-nitro-phenyl-beta-D-glucuronide. 3) The enzyme showed maximal activity at pH 5.0, compared with an optimum pH of 4.5 for beta-glucuronidase, which acts on p-nitro-phenyl-beta-D-glucuronide. 4) The enzyme showed maximal activity in 0.075 M NaCl but no activity above 0.25 M NaCl. 5) The enzyme was inhibited strongly by compounds bearing a sulfate group. 6) The enzyme did not react with an antibody against beta-glucuronidase acting on p-nitrophenyl-D-glucuronide. It is suggested that the enzyme may be involved in the catabolism of glycosaminoglycans, acting especially on chondroitin after the desulfation reaction and/or hyaluronic acid, but showing little involvement with the detoxification system. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|