Conversion of Red Fluorescent Protein into a Bright Blue Probe

Oksana M. Subach, Illia S. Gundorov, Masami Yoshimura, Fedor V. Subach, Jinghang Zhang, David Grüenwald, Ekaterina A. Souslova, Dmitriy M. Chudakov, Vladislav V. Verkhusha

Research output: Contribution to journalArticle

166 Scopus citations

Abstract

We used a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a rational design strategy to develop blue fluorescent proteins with a tyrosine-based chromophore. The strategy was applied to red fluorescent proteins of the different genetic backgrounds, such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which all were converted into blue probes. Further improvement of the blue variant of TagRFP by random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by the substantially higher brightness, the faster chromophore maturation, and the higher pH stability than blue fluorescent proteins with a histidine in the chromophore. The detailed biochemical and photochemical analysis indicates that mTagBFP is the true monomeric protein tag for multicolor and lifetime imaging, as well as the outstanding donor for green fluorescent proteins in Förster resonance energy transfer applications.

Original languageEnglish (US)
Pages (from-to)1116-1124
Number of pages9
JournalChemistry and Biology
Volume15
Issue number10
DOIs
StatePublished - Oct 20 2008

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Keywords

  • CHEMBIO

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Medicine
  • Molecular Biology
  • Pharmacology
  • Drug Discovery
  • Clinical Biochemistry

Cite this

Subach, O. M., Gundorov, I. S., Yoshimura, M., Subach, F. V., Zhang, J., Grüenwald, D., Souslova, E. A., Chudakov, D. M., & Verkhusha, V. V. (2008). Conversion of Red Fluorescent Protein into a Bright Blue Probe. Chemistry and Biology, 15(10), 1116-1124. https://doi.org/10.1016/j.chembiol.2008.08.006