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 journalArticlepeer-review

197 Citations (Scopus)


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
Pages (from-to)1116-1124
Number of pages9
JournalChemistry and Biology
Issue number10
Publication statusPublished - 20 Oct 2008
Externally publishedYes




Dive into the research topics of 'Conversion of Red Fluorescent Protein into a Bright Blue Probe'. Together they form a unique fingerprint.

Cite this