Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10765716 | Biochemical and Biophysical Research Communications | 2009 | 6 Pages |
Abstract
Blue fluorescent protein of aequorin (BFP) is a complex of Ca2+-bound apoaequorin with coelenteramide and is a bifunctional protein, which shows blue fluorescence and the luminescence activity like a luciferase. To reconstitute synthetic BFP (syn-BFP) from apoaequorin and coelenteramide, we established new synthetic route of coelenteramide and prepared highly purified recombinant aequorin using the histidine-tagged secretion system in Escherichia coli cells. As a result, we succeeded in reconstituting syn-BFP quantitatively and the fluorescence and luminescence properties of syn-BFP were identical to that of BFP obtained from aequorin.
Keywords
ApoaequorinGFPDTTRLUBFPSDS–PAGEEDTAEthylenediaminetetraacetic acidmatrix-assisted laser desorption/ionization time-of-flight mass spectrometrySodium dodecylsulfate-polyacrylamide gel electrophoresisBioluminescenceFluorescenceluciferaseMALDI-TOF-MSrelative light unitsBifunctional proteinCoelenterazine
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Biochemistry, Genetics and Molecular Biology
Biochemistry
Authors
Satoshi Inouye, Takamitsu Hosoya,