Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein

Hideaki Mizuno, Tapas Kumar Mal, Kit I. Tong, Ryoko Ando, Toshiaki Furuta, Mitsuhiko Ikura, Atsushi Miyawaki

Research output: Contribution to journalArticle

221 Citations (Scopus)

Abstract

Green fluorescent protein from the jellyfish (Aequorea GFP) and GFP-like proteins from coral species encode light-absorbing chromophores within their protein sequences. A coral fluorescent protein, Kaede, contains a tripeptide, His62-Tyr63-Gly64, which acts as a green chromophore that is photoconverted to red. Here, we present the structural basis for the green-to-red photoconversion. As in Aequorea GFP, a chromophore, 4-(p-hydroxybenzylidene)-5-imidazolinone, derived from the tripeptide mediates green fluorescence in Kaede. UV irradiation causes an unconventional cleavage within Kaede protein between the amide nitrogen and the α carbon (Cα) at His62 via a formal β-elimination reaction, which requires the whole, intact protein for its catalysis. The subsequent formation of a double bond between His62-Cα and -Cβ extends the π-conjugation to the imidazole ring of His62, creating a new red-emitting chromophore, 2-[(1E)-2-(5-imidazolyl)ethenyl]-4-(p- hydroxybenzylidene)-5-imidazolinone. The present study not only reveals diversity in the chemical structure of fluorescent proteins but also adds a new dimension to posttranslational modification mechanisms.

Original languageEnglish (US)
Pages (from-to)1051-1058
Number of pages8
JournalMolecular cell
Volume12
Issue number4
DOIs
StatePublished - Oct 2003

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Peptides
Anthozoa
Proteins
Post Translational Protein Processing
Green Fluorescent Proteins
Catalysis
Amides
Nitrogen
Carbon
Fluorescence
Light

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

Cite this

Mizuno, Hideaki ; Mal, Tapas Kumar ; Tong, Kit I. ; Ando, Ryoko ; Furuta, Toshiaki ; Ikura, Mitsuhiko ; Miyawaki, Atsushi. / Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein. In: Molecular cell. 2003 ; Vol. 12, No. 4. pp. 1051-1058.
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abstract = "Green fluorescent protein from the jellyfish (Aequorea GFP) and GFP-like proteins from coral species encode light-absorbing chromophores within their protein sequences. A coral fluorescent protein, Kaede, contains a tripeptide, His62-Tyr63-Gly64, which acts as a green chromophore that is photoconverted to red. Here, we present the structural basis for the green-to-red photoconversion. As in Aequorea GFP, a chromophore, 4-(p-hydroxybenzylidene)-5-imidazolinone, derived from the tripeptide mediates green fluorescence in Kaede. UV irradiation causes an unconventional cleavage within Kaede protein between the amide nitrogen and the α carbon (Cα) at His62 via a formal β-elimination reaction, which requires the whole, intact protein for its catalysis. The subsequent formation of a double bond between His62-Cα and -Cβ extends the π-conjugation to the imidazole ring of His62, creating a new red-emitting chromophore, 2-[(1E)-2-(5-imidazolyl)ethenyl]-4-(p- hydroxybenzylidene)-5-imidazolinone. The present study not only reveals diversity in the chemical structure of fluorescent proteins but also adds a new dimension to posttranslational modification mechanisms.",
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Mizuno, H, Mal, TK, Tong, KI, Ando, R, Furuta, T, Ikura, M & Miyawaki, A 2003, 'Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein', Molecular cell, vol. 12, no. 4, pp. 1051-1058. https://doi.org/10.1016/S1097-2765(03)00393-9

Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein. / Mizuno, Hideaki; Mal, Tapas Kumar; Tong, Kit I.; Ando, Ryoko; Furuta, Toshiaki; Ikura, Mitsuhiko; Miyawaki, Atsushi.

In: Molecular cell, Vol. 12, No. 4, 10.2003, p. 1051-1058.

Research output: Contribution to journalArticle

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T1 - Photo-induced peptide cleavage in the green-to-red conversion of a fluorescent protein

AU - Mizuno, Hideaki

AU - Mal, Tapas Kumar

AU - Tong, Kit I.

AU - Ando, Ryoko

AU - Furuta, Toshiaki

AU - Ikura, Mitsuhiko

AU - Miyawaki, Atsushi

PY - 2003/10

Y1 - 2003/10

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