The first mutant of the Aequorea victoria green fluorescent protein that forms a red chromophore

Alexander S. Mishin, Fedor V. Subach, Ilia V. Yampolsky, William King, Konstantin A. Lukyanov, Vladislav Verkhusha

Research output: Contribution to journalArticle

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Abstract

Green fluorescent protein (GFP) from a jellyfish, Aequorea victoria, and its mutants are widely used in biomedical studies as fluorescent markers. In spite of the enormous efforts of academia and industry toward generating its red fluorescent mutants, no GFP variants with emission maximum at more than 529 nm have been developed during the 15 years since its cloning. Here, we used a new strategy of molecular evolution aimed at generating a red-emitting mutant of GFP. As a result, we have succeeded in producing the first GFP mutant that substantially matures to the red-emitting state with excitation and emission maxima at 555 and 585 nm, respectively. A novel, nonoxidative mechanism for formation of the red chromophore in this mutant that includes a dehydration of the Ser65 side chain has been proposed. Model experiments showed that the novel dual-color GFP mutant with green and red emission is suitable for multicolor flow cytometry as an additional color since it is clearly separable from both green and red fluorescent tags.

Original languageEnglish (US)
Pages (from-to)4666-4673
Number of pages8
JournalBiochemistry
Volume47
Issue number16
DOIs
StatePublished - Apr 22 2008

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Chromophores
Green Fluorescent Proteins
Color
Molecular Evolution
Flow cytometry
Cloning
Dehydration
Organism Cloning
Industry
Flow Cytometry
Aequorea victoria green fluorescent protein
Experiments

ASJC Scopus subject areas

  • Biochemistry

Cite this

The first mutant of the Aequorea victoria green fluorescent protein that forms a red chromophore. / Mishin, Alexander S.; Subach, Fedor V.; Yampolsky, Ilia V.; King, William; Lukyanov, Konstantin A.; Verkhusha, Vladislav.

In: Biochemistry, Vol. 47, No. 16, 22.04.2008, p. 4666-4673.

Research output: Contribution to journalArticle

Mishin, AS, Subach, FV, Yampolsky, IV, King, W, Lukyanov, KA & Verkhusha, V 2008, 'The first mutant of the Aequorea victoria green fluorescent protein that forms a red chromophore', Biochemistry, vol. 47, no. 16, pp. 4666-4673. https://doi.org/10.1021/bi702130s
Mishin, Alexander S. ; Subach, Fedor V. ; Yampolsky, Ilia V. ; King, William ; Lukyanov, Konstantin A. ; Verkhusha, Vladislav. / The first mutant of the Aequorea victoria green fluorescent protein that forms a red chromophore. In: Biochemistry. 2008 ; Vol. 47, No. 16. pp. 4666-4673.
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