Chemical nature of the light emitter of the Aequorea green fluorescent protein.
The jellyfish Aequorea victoria possesses in the margin of its umbrella a green fluorescent protein (GFP, 27 kDa) that serves as the ultimate light emitter in the bioluminescence reaction of the animal. The protein is made up of 238 amino acid residues in a single polypeptide chain and produces a greenish fluorescence (lambda max = 508 nm) when irradiated with long ultraviolet light. The fluorescence is due to the presence of a chromophore consisting of an imidazolone ring, formed by a post-translational modification of the tripeptide -Ser65-Tyr66-Gly67-. GFP has been used extensively as a reporter protein for monitoring gene expression in eukaryotic and prokaryotic cells, but relatively little is known about the chemical mechanism by which fluorescence is produced. To obtain a better understanding of this problem, we studied a peptide fragment of GFP bearing the chromophore and a synthetic model compound of the chromophore. The results indicate that the GFP chromophore consists of an imidazolone ring structure and that the light emitter is the singlet excited state of the phenolate anion of the chromophore. Further, the light emission is highly dependent on the microenvironment around the chromophore and that inhibition of isomerization of the exo-methylene double bond of the chromophore accounts for its efficient light emission.
Niwa
H
H
Department of Applied Physics and Chemistry, University of Electro-Communications, Tokyo, Japan.
Inouye
S
S
Hirano
T
T
Matsuno
T
T
Kojima
S
S
Kubota
M
M
Ohashi
M
M
Tsuji
F I
FI
D000595:Amino Acid Sequence; D000818:Animals; D002863:Chromogenic Compounds / Q000138:chemical synthesis / Q000378:metabolism; D049452:Green Fluorescent Proteins; D007536:Isomerism; D008164:Luminescent Proteins / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000737:chemistry; D011499:Protein Processing, Post-Translational; D037401:Scyphozoa / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013053:Spectrophotometry
D002863:Chromogenic Compounds; D008164:Luminescent Proteins; D010446:Peptide Fragments; D049452:Green Fluorescent Proteins