Synthesis and properties of the red chromophore of the green-to-red photoconvertible fluorescent protein Kaede and its analogs

Ilia V. Yampolsky, Alexander A. Kislukhin, Tynchtyk T. Amatov, Dmitry Shcherbo, Victor K. Potapov, Sergey Lukyanov, Konstantin A. Lukyanov

Research output: Contribution to journalArticlepeer-review

48 Citations (Scopus)

Abstract

Green fluorescent protein (GFP) and homologous proteins possess a unique pathway of chromophore formation based on autocatalytic modification of their own amino acid residues. Green-to-red photoconvertible fluorescent protein Kaede carries His-Tyr-Gly chromophore-forming triad. Here, we describe synthesis of Kaede red chromophore (2-[(1E)-2-(5-imidazolyl)ethenyl]-4-(p-hydroxybenzylidene)-5-imidazolone) and its analogs that can be potentially formed by natural amino acid residues. Chromophores corresponding to the following tripeptides were obtained: His-Tyr-Gly, Trp-Tyr-Gly, Phe-Trp-Gly, Tyr-Trp-Gly, Asn-Tyr-Gly, Phe-Tyr-Gly, and Tyr-Tyr-Gly. In basic conditions they fluoresced red with relatively high quantum yield (up to 0.017 for Trp-derived compounds). The most red-shifted absorption peak at 595 nm was found for the chromophore Trp-Tyr-Gly in basic DMSO. Surprisingly, in basic DMF non-aromatic Asn-derived chromophore Asn-Tyr-Gly demonstrated the most red-shifted emission maximum at 642 nm. Thus, Asn residue may be a promising substituent, which can potentially diversify posttranslational chemistry in GFP-like proteins.

Original languageEnglish
Pages (from-to)96-104
Number of pages9
JournalBioorganic Chemistry
Volume36
Issue number2
DOIs
Publication statusPublished - Apr 2008
Externally publishedYes

Keywords

  • Bathochromic shift
  • Fluorescent probes
  • Fluorophore
  • Green fluorescent protein
  • Protein engineering

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