Reconstitution of blue fluorescent protein from recombinant apoaequorin and synthetic coelenteramide.
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.
Inouye
Satoshi
S
Yokohama Research Center, Chisso Corporation, 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan. sinouye@chisso.co.jp
Hosoya
Takamitsu
T
D000331:Aequorin / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D001059:Apoproteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D044482:Benzeneacetamides / Q000138:chemical synthesis / Q000737:chemistry*; D004926:Escherichia coli / Q000378:metabolism; D008164:Luminescent Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification; D011719:Pyrazines / Q000138:chemical synthesis / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000302:isolation & purification; D013050:Spectrometry, Fluorescence
D001059:Apoproteins; D044482:Benzeneacetamides; D008164:Luminescent Proteins; D011719:Pyrazines; D011994:Recombinant Proteins; C060014:apoaequorin; C409504:blue fluorescent protein, Aequorea victoria; C542661:coelenteramide; D000331:Aequorin