2020年2月26日
Expression, purification, and characterization of recombinant apoPholasin
Protein Expression and Purification
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- 巻
- 171
- 号
- 開始ページ
- 105615
- 終了ページ
- 105615
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1016/j.pep.2020.105615
Pholasin is a reactive oxygen-sensitive photoprotein that consists of an apoprotein (apoPholasin) and an unknown chromophore. The preferred human codon-optimized apoPholasin gene was transiently expressed in mammalian cells and apoPholasin was detected using an anti-recombinant apoPholasin antibody. For the first time, we found that apoPholasin secreted into the culture medium could catalyze the oxidation of coelenterazine (CTZ, a luciferin) to produce continuous luminescence. The fusion protein of apoPholasin and glutathione S-transferase (GST-apoPholasin) was successfully expressed as a soluble form in bacterial cells using the cold induction system. The purified GST-apoPholasin also had luminescence activity with CTZ, showing the bioluminescence emission peak at 461 nm, and the resultant product showed purple blue fluorescence under 365 nm light. Unexpectedly, the main oxidation product of CTZ was identified as coelenteramine (CTM), not coelenteramide (CTMD).
- リンク情報
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- DOI
- https://doi.org/10.1016/j.pep.2020.105615
- Web of Science
- https://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=JSTA_CEL&SrcApp=J_Gate_JST&DestLinkType=FullRecord&KeyUT=WOS:000525321400005&DestApp=WOS_CPL
- URL
- https://web.db.tokushima-u.ac.jp/cgi-bin/edb_browse?EID=363207
- URL
- https://www.ncbi.nlm.nih.gov/pubmed/32114101
- ID情報
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- DOI : 10.1016/j.pep.2020.105615
- ISSN : 1096-0279
- eISSN : 1096-0279
- Web of Science ID : WOS:000525321400005