| Project/Area Number |
05453211
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Functional biochemistry
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| Research Institution | Osaka University |
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Principal Investigator |
HASE Sumihiro Faculty of Science, Osaka University, Department of Chemistry, Professor, 理学部, 教授 (80028232)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥7,400,000 (Direct Cost: ¥7,400,000)
Fiscal Year 1994: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1993: ¥5,700,000 (Direct Cost: ¥5,700,000)
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| Keywords | mannosidase / sugar chain / cobalt / hen / pyridylamination / ピリジルアミノ化法 / 基質特異性 / 酵素 |
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| Research Abstract |
Several neutral alpha-mannosidases were reported in cytosol of cells. The mannosidases have unique substrate specificities after activation with Co(II). Howevere the only Purified enzyme was of quail oviduct, and the role of the enzyme and activation mechanism were unknown.
alpha-Mannosidase activated with Co(II) was purified from hen oviduct to homogeneity as judged by native and SDS PAGE.Its substrate specificity was analyzed by using pyridylaminated sugar chains. The mannosidase was found to belong to the neutral alpha-mannosidase in cytosol fraction. The results show that substrate specificities and activation with Co(II) of the alpha-mannosidase from hen oviduct were the same as those of quail oviduct. Analysis of the optimal activation pH,optimal concentration of Co(II), protease activity by analysis with several peptides, proteinase inhibitors indicate that the mannosidase contains protease. However, the activated mannosidase showed almost the same electrophoretic patterns as compared with the one without activation. Further analysis of the structure of the enzyme is needed to clarify the exact mechanism.
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