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昆虫細胞を利用したカテプシンK酵素の作製
https://doi.org/10.15069/00000908
https://doi.org/10.15069/000009085f033ff3-bc3d-41f1-aa7a-90f3af9e196e
名前 / ファイル | ライセンス | アクション |
---|---|---|
31 研究紀要 10号 定金 (467.9 kB)
|
Item type | 紀要論文(ELS) / Departmental Bulletin Paper(1) | |||||
---|---|---|---|---|---|---|
公開日 | 2009-03-25 | |||||
タイトル | ||||||
タイトル | 昆虫細胞を利用したカテプシンK酵素の作製 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Preparation of recombinant lysosomal cysteine protease, cathepsin K in insect cells | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | 昆虫細胞 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | 組換え酵素 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | 分泌型酵素 | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | mycタグ | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | insect cell | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | recombinant enzyme | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | secretory enzyme | |||||
キーワード | ||||||
言語 | en | |||||
主題Scheme | Other | |||||
主題 | myc-tag | |||||
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 | |||||
ID登録 | ||||||
ID登録 | 10.15069/00000908 | |||||
ID登録タイプ | JaLC | |||||
ページ属性 | ||||||
内容記述タイプ | Other | |||||
内容記述 | P(論文) | |||||
著者名(日) |
定金, 豊
× 定金, 豊× 伊藤, 武朗× 今西, 重雄× 中込, 和哉× 畑中, 保丸 |
|||||
著者名よみ |
サダカネ, ユタカ
× サダカネ, ユタカ× イトウ, タケアキ× イマニシ, シゲオ× ナカゴミ, カズヤ× ハタナカ, ヤスマル |
|||||
著者名(英) |
SADAKANE, Yutaka
× SADAKANE, Yutaka× ITO, Takeaki× IMANISHI, Shigeo× NAKAGOMI, Kazuya× HATANAKA, Yasumaru |
|||||
著者所属(日) | ||||||
値 | 九州保健福祉大学薬学部薬学科 | |||||
著者所属(日) | ||||||
値 | 富山医科薬科大学薬学部 | |||||
著者所属(日) | ||||||
値 | 農業生物資源研究所昆虫生産工学研究グループ | |||||
著者所属(日) | ||||||
値 | 帝京大学薬学部 | |||||
著者所属(日) | ||||||
値 | 富山医科薬科大学薬学部 | |||||
著者所属(英) | ||||||
言語 | en | |||||
値 | Department of Pharmaceutical Sciences, School of Pharmaceutical Sciences, Kyushu University of Health and Welfare | |||||
著者所属(英) | ||||||
言語 | en | |||||
値 | Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical Univesity | |||||
著者所属(英) | ||||||
言語 | en | |||||
値 | Insect Biotechnology and Sericology Department, The National Institute of Agrobiological Sciences | |||||
著者所属(英) | ||||||
言語 | en | |||||
値 | School of Pharmaceutical Sciences, Teikyo University | |||||
著者所属(英) | ||||||
言語 | en | |||||
値 | Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical Univesity | |||||
抄録(日) | ||||||
内容記述タイプ | Other | |||||
内容記述 | 軟骨細胞に局在するシステインプロテアーゼであるカテプシンKは,骨の差異九州に関わる酵素である。カテプシンKの阻害剤は骨粗鬆症の治療薬の候補になり得る。スクリーニングや酵素の機能を調べるためには組み換えカテプシンKを大量に作成することが必要である。我々はカイコ幼虫を用いた発現系が分泌型酵素であるカテプシンKの発現に適していると考えた。C末端にmycタグを付けたカテプシンK遺伝子をカイコ用の発現ベクターに挿入し,限界希釈法で組み換えウイルスを得た。このウイルスをカイコ培養細胞に感染させてカテプシンKの発現場所を調べたところ,細胞内で発現していた。細胞外への分泌が観察されなかったのでカイコ幼虫での発現をあきらめ,培養細胞からの精製を試みた。DEAE-セファロースカラム,Mono-Sカラム,抗myc抗体アフィニティーカラムで順次精製した結果,非活性を細胞可溶性画分の約300倍に高めることに成功した。 | |||||
抄録(英) | ||||||
内容記述タイプ | Other | |||||
内容記述 | Cathepsin K, which is localized in osteoclasts, plays an important role of bone resorption. Selective inhibitors of cathepsin K therefore can be promising therapeutic candidates for the treatment of diseases such as osteoporosis. Recombinant cathepsin K protein is necessary for screening the therapeutic candidates and elucidating the enzymatic functions because the amount of native cathepsin K is very low. Silkworm is one of the most attractive hosts for large-scale production of eukaryotic secretory proteins. The gene for fusion cathepsin K protein with myc-tag at the C-terminus was inserted into expression vector for silkworm and recombinant viruses were isolated by end-point dilution assay. Cathepsin K was expressed in the cell lysate, but not in the medium, and consequently we gave up the idea of cathepsin K expression in the silkworm. Thus, we tried to isolate cathepsin K protein from the cultured cells, and purified it by sequential operations with DEAE-Sepharose column, Mono-S column, and anti-myc antibody column. The purification fold of cathepsin K activity increased approximately 300 times in comparison with that in soluble fraction of cell lysate. | |||||
雑誌書誌ID | ||||||
収録物識別子タイプ | NCID | |||||
収録物識別子 | AA11490417 | |||||
書誌情報 |
九州保健福祉大学研究紀要 en : Journal of Kyushu University of Health and Welfare 巻 10, p. 201-207, 発行日 2009-03 |