Abstract
Blue fluorescent protein of aequorin (BFP) is a complex of Ca(2+)-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.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Aequorin / biosynthesis*
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Aequorin / chemistry*
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Aequorin / isolation & purification
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Apoproteins / biosynthesis*
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Apoproteins / chemistry*
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Apoproteins / isolation & purification
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Benzeneacetamides / chemical synthesis
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Benzeneacetamides / chemistry*
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Escherichia coli / metabolism
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Luminescent Proteins / biosynthesis*
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Luminescent Proteins / chemistry*
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Luminescent Proteins / isolation & purification
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Pyrazines / chemical synthesis
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Pyrazines / chemistry*
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Recombinant Proteins / biosynthesis
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Recombinant Proteins / chemistry
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Recombinant Proteins / isolation & purification
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Spectrometry, Fluorescence
Substances
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Apoproteins
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Benzeneacetamides
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Luminescent Proteins
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Pyrazines
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Recombinant Proteins
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apoaequorin
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blue fluorescent protein, Aequorea victoria
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coelenteramide
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Aequorin