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Production of Human Type Glycosylated Tissue Plasminogen Activator and the Role of Its Carbohydrate Moiety
https://tokushima-u.repo.nii.ac.jp/records/2001118
https://tokushima-u.repo.nii.ac.jp/records/200111898c50532-e012-499c-af8b-35adaf79cc8c
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
EID185295.pdf (880 KB)
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| Item type | 文献 / Documents(1) | |||||||||||||||||||||||
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| 公開日 | 2011-08-11 | |||||||||||||||||||||||
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| アクセス権 | open access | |||||||||||||||||||||||
| 資源タイプ | ||||||||||||||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||||||||||||||
| 資源タイプ | departmental bulletin paper | |||||||||||||||||||||||
| 出版タイプ | ||||||||||||||||||||||||
| 出版タイプ | VoR | |||||||||||||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||||||||||||||
| タイトル | ||||||||||||||||||||||||
| タイトル | Production of Human Type Glycosylated Tissue Plasminogen Activator and the Role of Its Carbohydrate Moiety | |||||||||||||||||||||||
| 言語 | en | |||||||||||||||||||||||
| 著者 |
三木, 瑞枝
× 三木, 瑞枝
× 谷本, 久美
× 四方, 知子
× 横井川, 久己男 |
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| 抄録 | ||||||||||||||||||||||||
| 内容記述タイプ | Abstract | |||||||||||||||||||||||
| 内容記述 | To begin the examination of the role of human type carbohydrate moiety of tissue plasminogen activator (t-PA) on the binding of the enzyme to fibrin，the naturally glycosylated enzyme was produced by microcarrier culture of human cells established from normal uterine muscle. The cells grown on microcarrers in Haunks' MEM supplemented with 10 % FBS (1.6 x 10 6 cells/ml) rapidly detached themselves from microcarrier in a serum-free medium (t-PA production medium) within 5 days, and it was difficult to produce t-PA for long time (t-PA production: an average of 3 IU/ml/day over 5 days). Addition of 0.5% beef extract to the serum-free medium suppressed their detaching from microcarriers. By regulating the pH (7.4) and dissolved oxygen(4 ppm) of the serum free medium，the cell density of microcarrier culture increased to 1.2 x10 7 cells/ml and t-PA was produced over 38 days (t-PA production: an average of 836 IU/ml/day over 38 days). Native t-PAs purified to homogeneity from the culture broth had the molecular weight of 63,000 and 65,000 containing 5.6 and 8.5 % carbohydrate，respectively (molecular mass of protein moiety was calculated to be 59,500). By enzymatic digestion of carbohydrate moiety in native t-PA，we obtained partially deglycosylated t-PA with molecular weights of 60,000 and 62,000. Completely deglycosylated t-PA was obtained by t-PA production in the presence of 10μg/ml tunicamycin (N-glycosylation inhibitor). This suggests that N-glycosylationof t-PA occurs while O-glycosylation is absent. The binding strength of these enzymes to fibrin increased with decrease of the carbohydrate content. The carbohydrate moiety of human type glycosylated t-PA probably modulates the binding strength of the enzyme to fibrin. |
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| 言語 | en | |||||||||||||||||||||||
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| 言語 | en | |||||||||||||||||||||||
| 主題Scheme | Other | |||||||||||||||||||||||
| 主題 | Tissue plasminogen activator | |||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||
| 言語 | en | |||||||||||||||||||||||
| 主題Scheme | Other | |||||||||||||||||||||||
| 主題 | cell culture | |||||||||||||||||||||||
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| 言語 | en | |||||||||||||||||||||||
| 主題Scheme | Other | |||||||||||||||||||||||
| 主題 | carbohydrate | |||||||||||||||||||||||
| キーワード | ||||||||||||||||||||||||
| 言語 | en | |||||||||||||||||||||||
| 主題Scheme | Other | |||||||||||||||||||||||
| 主題 | animal cell | |||||||||||||||||||||||
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| 言語 | en | |||||||||||||||||||||||
| 主題Scheme | Other | |||||||||||||||||||||||
| 主題 | fibrinolysis | |||||||||||||||||||||||
| bibliographic_information |
ja : 徳島大学総合科学部自然科学研究 = Natural Science Research, The University of Tokushima 巻 22, p. 97-104, 発行日 2008-12 |
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| 収録物識別子タイプ | ISSN | |||||||||||||||||||||||
| 収録物識別子 | 09146385 | |||||||||||||||||||||||
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| 収録物識別子タイプ | NCID | |||||||||||||||||||||||
| 収録物識別子 | AN10065859 | |||||||||||||||||||||||
| 出版者 | ||||||||||||||||||||||||
| 出版者 | 徳島大学総合科学部 | |||||||||||||||||||||||
| 言語 | ja | |||||||||||||||||||||||
| item_1723180141928 | ||||||||||||||||||||||||
| 識別子 | 185295 | |||||||||||||||||||||||
| 識別子タイプ | URI | |||||||||||||||||||||||
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| 言語 | eng | |||||||||||||||||||||||
