| Project/Area Number |
62580112
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | Ochanomizu University |
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Principal Investigator |
SENO Nobuko Professor of Ochanomizu University, 理学部, 教授 (40017182)
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| Co-Investigator(Kenkyū-buntansha) |
K OGAWA Haruko Instructor of Ochanomizu University, 理学部, 助手 (90143700)
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| Project Period (FY) |
1987 – 1988
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| Project Status |
Completed (Fiscal Year 1988)
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| Budget Amount *help |
¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1988: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 1987: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | -Glucuronidase / Lysosomal enzyme / Affinity chromatography / ラクタミルSepharose / β-グルクロニダーゼ / ラクタミルSepharose. / アフィニティクロマトグラフィー |
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| Research Abstract |
Bovine liver -glucuronidase is one of the important lysosomal enzymes degrading glycosaminoglycans. The chemical and enzymatic properties have been investigated, while the regulation mechanism of the enzyme activity in vivo has not yet been elucidated. In this study, the effects of various substances on the activities of -glucuronidases from different sources were extensively investigated. Among the various saccharides tested, lactose was found to be the best activator for -glucuronidase from bovine and squid livers but not from E. coli, while saccharo-1,4-lactone was a typical competitive inhibitor for all of the enzymes. In addition, it was found that the activity of bovine -glucuronidase was affected by various concentrations of thyroglobulin and asialo-thyroglobulin, but not by nonglycosylated protein. These results suggested that the saccharides or saccharide moieties of glycoproteins regulate the activities of lysosomal -glucuronidases, although further studies are required. On the basis of above results, commercial bovine -glucuronidase contaminated with N-acetylhexosaminidases was successfully purified by affinity chromatography on lactamyl-Sepharose. The recovery of the enzyme activity was over 70 % and the purified enzyme showed a single band on SDS-PAGE.
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