Identification of a novel oxidation product from yellow fluorophore in the complex of apoPholasin and dehydrocoelenterazine. Issue 19 (1st October 2020)

• Record Type: Journal Article
• Authors: Inouye, Satoshi; Nakamura, Mitsuhiro; Taguchi, Jumpei; Hosoya, Takamitsu
• Is Part Of: Bioorganic & medicinal chemistry letters. Volume 30:Issue 19(2020)
• Publication Date: 2020-10-01
• Subjects: apoPholasin apoprotein of pholasin -- CTZ coelenterazine -- dCTZ dehydrocoelenterazine -- CTMD coelenteramide -- CTM coelenteramine -- CTO 3-benzyl-5-(4-hydroxyphenyl)pyrazin-2(1H)-one -- GST glutathione S-transferase -- GST-apoPholasin the fusion protein of GST and apoPholasin -- GST-apoPholasin/dCTZ complex a complex of GST-apoPholasin and dCTZ -- dCTZ* fluorescent chromophore in GST-apoPholasin/dCTZ complex -- Imax maximum luminescence intensity -- rlu relative light units; Coelenterazine -- Coelenteramide -- Coelenteramine -- Photoproteins -- Reactive oxygen; Bioorganic chemistry -- Periodicals; Pharmaceutical chemistry -- Periodicals; 572
• DOI: 10.1016/j.bmcl.2020.127435
• Languages: English
• ISSNs: 0960-894X

Abstract

Graphical abstract: Highlights: The apoPholasin/dCTZ complex shows considerably weak continuous luminescence. The complex converts to blue fluorescent protein spontaneously. The oxidation products are coelenteramine (CTM) and a novel pyrazine compound (CTO). Chemical synthesized CTO was used for identification by mass spectrometry. CTM and CTO likely originate form imine intermediates of excited CTMD. Abstract: The complex of the recombinant fusion protein of apoPholasin and glutathione S -transferase (GST-apoPholasin) with non-fluorescent dehydrocoelenterazine (dCTZ) (GST-apoPholasin/dCTZ complex) shows yellow fluorescence at 539 nm by excitation at 430 nm. The GST-apoPholasin/dCTZ complex with a fluorophore (dCTZ*) has considerably weak luminescence activity, converting slowly to a blue fluorescence protein with the emission peak at 430 nm. The main oxidation products from dCTZ* for blue fluorescence were identified as coelenteramine (CTM) and an unreported pyrazine derivative, 3-benzyl-5-(4-hydroxyphenyl)pyrazin-2(1 H )-one (CTO) that was confirmed by chemical synthesis.