2007年4月
Gliadain, a gibberellin-inducible cysteine proteinase occurring in germinating seeds of wheat, Triticum aestivum L., specifically digests gliadin and is regulated by intrinsic cystatins
FEBS JOURNAL
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- 巻
- 274
- 号
- 8
- 開始ページ
- 1908
- 終了ページ
- 1917
- 記述言語
- 英語
- 掲載種別
- DOI
- 10.1111/j.1742-4658.2007.05749.x
- 出版者・発行元
- BLACKWELL PUBLISHING
We cloned a new cysteine proteinase of wheat seed origin, which hydrolyzed the storage protein gliadin almost specifically, and was named gliadain. Gliadain mRNA was expressed 1 day after the start of seed imbibition, and showed a gradual increase thereafter. Gliadain expression was suppressed when uniconazol, a gibberellin synthesis inhibitor, was added to germinating seeds. Histochemical detection with anti-gliadain serum indicated that gliadain was present in the aleurone layer and also that its expression intensity increased in sites nearer the embryo. The enzymological characteristics of gliadain were investigated using recombinant glutathione S-transferase (GST)-progliadain fusion protein produced in Escherichia coli. The GST-progliadain almost specifically digested gliadin into low molecular mass peptides. These results indicate that gliadain is produced via gibberellin-mediated gene activation in aleurone cells and secreted into the endosperm to digest its storage proteins. Enzymologically, the GST-progliadain hydrolyzed benzyloxycarbonyl-Phe-Arg-7-amino-4-methylcoumarin (Z-Phe-Arg-NH2-Mec) at K-m = 9.5 mu M, which is equivalent to the K-m value for hydrolysis of this substrate by cathepsin L. Hydrolysis was inhibited by two wheat cystatins, WC1 and WC4, with IC50 values of 1.7 x 10(-8) and 5.0 x 10(-8) M, respectively. These values are comparable with those found for GST-progliadain inhibition by E-64 and egg-white cystatin, and are consistent with the possibility that, in germinating wheat seeds, gliadain is under the control of intrinsic cystatins.
- リンク情報
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- DOI
- https://doi.org/10.1111/j.1742-4658.2007.05749.x
- CiNii Articles
- http://ci.nii.ac.jp/naid/80018607838
- PubMed
- https://www.ncbi.nlm.nih.gov/pubmed/17371549
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000245506600003&DestApp=WOS_CPL
- ID情報
-
- DOI : 10.1111/j.1742-4658.2007.05749.x
- ISSN : 1742-464X
- CiNii Articles ID : 80018607838
- PubMed ID : 17371549
- Web of Science ID : WOS:000245506600003