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Blue fluorescent protein from the calcium‐sensitive photoprotein aequorin is a heat resistant enzyme, catalyzing the oxidation of coelenterazine
Author(s) -
Inouye Satoshi
Publication year - 2004
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/j.febslet.2004.09.078
Subject(s) - aequorin , photoprotein , bioluminescence , chemistry , fluorescence , luciferin , ethylenediaminetetraacetic acid , green fluorescent protein , luciferase , biochemistry , photochemistry , inorganic chemistry , chelation , transfection , physics , quantum mechanics , gene , intracellular
Blue fluorescent protein from the calcium‐sensitive photoprotein aequorin (BFP‐aq) was prepared and determined to be a heat resistant enzyme, catalyzing the luminescent oxidation of coelenterazine (luciferin) with molecular oxygen as a general luciferase. After treatment with excess ethylenediaminetetraacetic acid to remove Ca 2+ from BFP‐aq, the blue fluorescence shifted to a greenish fluorescence. This greenish fluorescent protein (gFP‐aq) was identified as a non‐covalent complex of apoaequorin with coelenteramide (oxyluciferin) in a molar ratio of 1:1. By incubation with coelenterazine in the absence of reducing reagents, gFP‐aq was converted to aequorin at 25 °C. BFP‐aq and gFP‐aq possessing both fluorescence and luminescence activities may work as novel reporter proteins.