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Identification of Cysteine Proteases and Screening of Cysteine Protease Inhibitors in Biological Samples by a Two-Dimensional Gel System of Zymography and Reverse Zymography
http://hdl.handle.net/10191/30169
http://hdl.handle.net/10191/30169b4924d44-b3b2-4a85-a712-03df2fa29a3e
名前 / ファイル | ライセンス | アクション |
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2_51-59.pdf (1.4 MB)
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Item type | 学術雑誌論文 / Journal Article(1) | |||||
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公開日 | 2014-11-12 | |||||
タイトル | ||||||
言語 | en | |||||
タイトル | Identification of Cysteine Proteases and Screening of Cysteine Protease Inhibitors in Biological Samples by a Two-Dimensional Gel System of Zymography and Reverse Zymography | |||||
言語 | ||||||
言語 | eng | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Broad bean lectin | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Cysteine proteases | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Cysteine protease inhibitors | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Reverse zymography | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Two-dimensional gel electrophoresis | |||||
キーワード | ||||||
主題Scheme | Other | |||||
主題 | Zymography | |||||
資源タイプ | ||||||
資源 | http://purl.org/coar/resource_type/c_6501 | |||||
タイプ | journal article | |||||
著者 |
Saitoh, Eiichi
× Saitoh, Eiichi× Yamamoto, Shinya× Okamoto, Eishiro× Hayakawa, Yoshimi× Hoshino, Takashi× Sato, Ritsuko× Isemura, Satoko× Ohtsubo, Sadami× Taniguchi, Masayuki |
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抄録 | ||||||
内容記述タイプ | Abstract | |||||
内容記述 | We have developed a two-dimensional (2D-) gel system of zymography and reverse zymography for the detection and characterization of proteases and protease inhibitors. Isoelectric focusing (IEF) agarose gels with pH gradients were employed for separation in the fi rst-dimension and sodium dodecyl sulfate (SDS)-polyacrylamide gel copolymerized with gelatin used for the second dimension. Proteases and protease inhibitors separated by IEF gel were applied on the second gel without trichloroacetic acid (TCA) fi xation. Protease activity in the 2D-gel was visualized as transparent spots where gelatin substrate was digested after commassie brilliant blue (CBB) staining. Some of the transparent spots from the skin mucus extract of rainbow trout were determined to be a cysteine protease through use of E-64 or CA-074. In the reverse zymography technique, the gel was incubated with papain solution at 37 ºC for 18 h. Cysteine protease inhibitors from broad bean seeds were detected as clear blue spots after CBB staining. The amino (N-) terminal sequences of four papain inhibitor spots thus detected were demonstrated to be identical to that of favin β chain, a broad bean lectin. Taken together, our system can be considered to be an effi cient technique for discovering and characterizing new proteases and protease inhibitors in biological samples. This is the fi rst report describing a 2D-gel system of zymography and reverse zymography. | |||||
書誌情報 |
en : Analytical Chemistry Insights 巻 2, p. 51-59, 発行日 2007-11 |
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出版者 | ||||||
出版者 | Libertas Academica | |||||
ISSN | ||||||
収録物識別子タイプ | ISSN | |||||
収録物識別子 | 1177-3901 | |||||
DOI | ||||||
識別子タイプ | DOI | |||||
関連識別子 | http://doi.org/10.4137/117739010700200011 | |||||
権利 | ||||||
権利情報 | Copyright(C) 2007 Libertas Academica | |||||
出版タイプ | ||||||
出版タイプ | VoR | |||||
出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||
著者版フラグ | ||||||
値 | publisher |