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Conversion of Red Fluorescent Protein into a Bright Blue Probe
Open accessAbstract
SUMMARY We utilized a red chromophore formation pathway, in which the anionic red chromophore is formed from the neutral blue intermediate, to suggest a novel 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 background such as TagRFP, mCherry, HcRed1, M355NA, and mKeima, which were converted into blue probes. Further improvement of a blue variant of TagRFP using random mutagenesis resulted in an enhanced monomeric protein, mTagBFP, characterized by substantially higher brightness, faster chromophore maturation and higher pH stability than blue fluorescence proteins with a histidine in the chromophore. Detailed biochemical and photochemical analysis indicates mTagBFP is the true monomeric protein tag for multicolor and lifetime imaging as well as the outstanding donor for green fluorescent proteins in FRET applications.
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Shaner, N. C., Campbell, R. E., Steinbach, P. A., Giepmans, B. N. G., Palmer, A. E., & Tsien, R. Y. (2004) · Nature Biotechnology · doi:10.1038/nbt1037 · Cited by 4,619
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Self-citationKoike-Yusa, H., Li, Y., Tan, E.-P., Velasco-Herrera, M. D. C., & Yusa, K. (2014) · Nature Biotechnology · doi:10.1038/nbt.2800 · Cited by 1,064
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Metadata
- Authors
- Subach, O. M., Gundorov, I. S., Yoshimura, M., Subach, F. V., Zhang, J., Grüenwald, D., Souslova, E. A., Chudakov, D. M., & Verkhusha, V. V.
- Venue
- Chemistry & Biology
- Publisher
- Elsevier BV
- Published
- 2008-10
- Volume
- 15
- Issue
- 10
- Pages
- 1116-1124
- ISSN
- 1074-5521 (ISSN-L)
- OpenAlex
- W2053679951
- Cited by
- 303
- References
- 1
- FWCI
- 6.23
- Type
- journal article
- Language
- en
- Open access
- bronze
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- 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 & Biology. https://doi.org/10.1016/j.chembiol.2008.08.006
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