Blue fluorescent protein from the calcium-sensitive photoprotein aequorin: catalytic properties for the oxidation of coelenterazine as an oxygenase.
Blue fluorescent protein from the calcium-binding photoprotein aequorin (BFP-aq) is a complex of Ca2+ -bound apoaequorin and coelenteramide, and shows luminescence activity like a luciferase, catalyzing the oxidation of coelenterazine with molecular oxygen. To understand the catalytic properties of BFP-aq, various fluorescent proteins (FP-aq) have been prepared from semi-synthetic aequorin and characterized in comparison with BFP-aq. FP-aq has luciferase activity and could be regenerated into native aequorin by incubation with coelenterazine. The results from substrate specificity studies of FP-aq using various coelenterazine analogues have suggested that the oxidation of coelenterazine by BFP-aq in the luciferase reaction and the regeneration process to aequorin might involve the same catalytic site of BFP-aq.
Inouye
Satoshi
S
Yokohama Research Center, Chisso Co., 5-1 Okawa, Kanazawa-ku, Yokohama 236-8605, Japan. sinouye@chisso.co.jp
Sasaki
Satoko
S
D000331:Aequorin / Q000378:metabolism*; D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002384:Catalysis; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism*; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D008164:Luminescent Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D010084:Oxidation-Reduction; D010105:Oxygenases / Q000378:metabolism*; D011719:Pyrazines / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000378:metabolism; D013050:Spectrometry, Fluorescence; D013379:Substrate Specificity
D007093:Imidazoles; D008164:Luminescent Proteins; D011719:Pyrazines; D011994:Recombinant Proteins; C409504:blue fluorescent protein, Aequorea victoria; C017144:coelenterazine; D000331:Aequorin; D010105:Oxygenases; D008156:Luciferases; D002118:Calcium