Intermolecular energy transfer in the bioluminescent system of Aequorea

Morise, Hiroshi; Shimomura, Osamu; Johnson, Frank H.; Winant, John

doi:10.1021/bi00709a028

Cited by 406 publications

(290 citation statements)

References 29 publications

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“…3). The observed maxima and shapes of the spectra were virtually the same as to those that have been previously reported for native GFP [6,26]. The fluorescence of native GFP has been attributed to the presence of an hexapeptide chromophore (Fig.…”

Section: Resultssupporting

confidence: 83%

“…Since native GFP is known to self-associate [6,27], the 43 kDa band is most likely due to the presence of a partially denatured GFP dimer, which is converted to a monomer (31 kDa) by denaturation in lane 2. This would account for the bright greenish fluorescence of the 43 kDa band in lane 3 [6], and for its absence in lane 4 as a fluorescent band and in lane 2 as a stained band. On denaturation, GFP is known to become completely non-fluorescent [26,28].…”

Section: Resultsmentioning

confidence: 99%

“…The blue fluorescent protein is made up of coelenteramide bound to apoaequorin and the excited state coelenteramide is the emitter in the reaction. If GFP is added to the reaction mixture, however, a bluish-green light (&,, = 508 nm), identical to that of the in vivo luminescence of the animal, is observed due to a radiationless energy transfer from the blue fluorescent protein to GFP [5,6,11]. The green light emission may be schematically outlined, as follows: The genes for apoaequorin and GFP have been cloned and the primary structure of the proteins deduced from the nucleotide sequence [3,12].…”

Section: Introductionmentioning

confidence: 99%

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Aequorea green fluorescent protein

1994

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Expression of the cDNA for Aequorea green fluorescent protein in E. coli yielded a fused protein with fluorescence excitation and emission spectra virtually identical to those of the native green fluorescent protein. Further, a solution of the protein, when mixed with aequorin and calcium ion, emitted a greenish luminescence characteristic of the in vivo luminescence of the animal, indicating a radiationless energy transfer to the protein.

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Section: Resultssupporting

confidence: 83%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Aequorea green fluorescent protein

1994

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“…Regarding its photophysics, the wild type GFP protein is characterized by high quantum yields, ϕ f~0 .8, in contrast to what is observed for its individual cromophore in aqueous solution, ϕ f~1 0 −3 [14,15]. In the native fold, the inhibition of molecular vibration or rotation along the chromophore double exo-metilene bond, prevents emission quenching by a rapid internal conversion process [13,15,16].…”

Section: Introductionmentioning

confidence: 98%

Thermal Effect on Aequorea Green Fluorescent Protein Anionic and Neutral Chromophore Forms Fluorescence

Santos

2011

J Fluoresc

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The emission behaviour of Aequorea green fluorescent protein (A-GFP) chromophore, in both neutral (N) and anionic (A) form, was studied in the temperature range from 20°C to 75°C and at pH=7. Excitation wavelengths of 399 nm and 476 nm were applied to probe the N and A forms environment, respectively. Both forms exhibit distinct fluorescence patterns at high temperature values. The emission quenching rate, following a temperature increase, is higher for the chromophore N form as a result of the hydrogen bond network weakening. The chromophore anionic form emission maximum is red shifted, upon temperature increase, due to a charge transfer process occurring after A form excitation.

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“…No additive (calcium, a prosthetic group) is necessary for Aequorin to emit light 4,8,9,12,14,15,16 .…”

Section: Introductionmentioning

confidence: 99%

Genótipo e expressão da proteína fluorescente verde melhorada em ratos da linhagem LEW-Tg (EGFP) F455.5/Rrrc

Brito

Carvalho

Zonari

et al. 2013

Braz. J. Vet. Res. Anim. Sci.

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Genotype and expression of the enhanced green fluorescent protein in LEW-Tg (EGFP) F455.5/Rrrc rats AbstractThe Green fluorescent protein (GFP) was first described after being extracted from Aequorea victoria in 1987; Since then, GFP and its derivatives have been widely used in several experiments as cell and protein marker. In the present study it was verified the genotype of the offspring from crosses between heterozygote Lewis LEW-Tg (EGFP) F455.5/ Rrrc rats and analyzed the expression of the enhanced green fluorescent protein (EGFP) in different cell types and genotypes. The genotype of the offspring was assessed by PCR and analysis of EGFP expression in different cells and genotypes, including mesenchymal stem cells (MSC) derived from adipose tissue and calvarial osteoblast cells. Expression of EGFP was verified by flow cytometry, fluorescence microscopy, and immunostaining. Through these methods, it was identified the genotypes of the offspring and determined the levels of expression of EGFP in two cell types. A difference in expression between the (EGFP +/+) and (EGFP +/-) genotypes was also observed in addition to the presence of autofluorescence. Further studies on the natural fluorescence of cells with the (EGFP +/-) genotype and that induced by presence of the EGFP are necessary. Keywords: Enhanced green fluorescent protein. EGFP. Autofluorescence. Difference in expression. ResumoA proteína fluorescente verde (GFP) foi descrita pela primeira vez após ter sido extraída de Aequorea victoria em 1987. Desde então, a GFP e seus derivados têm sido amplamente utilizados em várias experiências como marcador celular e de proteínas. O objetivo do presente estudo foi o de verificar o genótipo dos descendentes de cruzamentos entre ratos Lewis LEW-Tg (EGFP) F455.5/Rrrc heterozigotos e de analisar a expressão da proteína fluorescente verde melhorada (EGFP) em diferentes tipos celulares e genótipos. O genótipo da descendência foi avaliado por PCR e pela análise da expressão da EGFP em diferentes células e genótipos, incluindo-se as células-tronco mesenquimais (MSC) derivadas de tecido adiposo e de osteoblastos de calvária. A expressão da EGFP foi verificada por citometria de fluxo, microscopia de fluorescência e imunocoloração. Foram, identificados os genótipos da descendência e determinados os níveis de expressão de EGFP em dois tipos de células. Foi também constatada uma diferença de expressão entre os genótipos (EGFP +/+) e (EGFP +/-) além da presença de autofluorescência. Mais estudos são necessários para esclarecer a fluorescência natural de células com o genótipo (EGFP +/-) e aquela induzida pela presença da EGFP.Palavras-chave: Proteína verde fluorescente melhorada. (EGFP). Autofluorescência. Diferença de expressão.

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Intermolecular energy transfer in the bioluminescent system of Aequorea

Cited by 406 publications

References 29 publications

Aequorea green fluorescent protein

Aequorea green fluorescent protein

Thermal Effect on Aequorea Green Fluorescent Protein Anionic and Neutral Chromophore Forms Fluorescence

Genótipo e expressão da proteína fluorescente verde melhorada em ratos da linhagem LEW-Tg (EGFP) F455.5/Rrrc

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