An enhanced monomeric blue fluorescent protein with the high chemical stability of the chromophore

Oksana M. Subach, Paula J. Cranfill, Michael W. Davidson, Vladislav Verkhusha

Research output: Contribution to journalArticle

104 Citations (Scopus)

Abstract

Commonly used monomeric blue fluorescent proteins suffer from moderate brightness. The brightest of them, mTagBFP, has a notably low chemical stability over time. Prolonged incubation of mTagBFP leads to its transition from a blue fluorescent state with absorbance at 401 nm to a non-fluorescent state with absorbance at 330 nm. Here, we have determined the chemical structure of the degraded product of the blue mTagBFP-like chromophore. On the basis of mTagBFP we have developed an improved variant, named mTagBFP2. mTagBFP2 exhibits 2-fold greater chemical stability and substantially higher brightness in live cells than mTagBFP. mTagBFP2 is also 1.2-fold and 1.7-fold more photostable than mTagBFP in widefield and confocal microscopy setups, respectively. mTagBFP2 maintains all other beneficial properties of the parental mTagBFP including the high pH stability and fast chromophore formation. The enhanced photostability and chromophore chemical stability of mTagBFP2 make it a superior protein tag. mTagBFP2 performs well in the numerous protein fusions and surpasses mTagBFP as a donor in Förster resonance energy transfer with several green fluorescent protein acceptors.

Original languageEnglish (US)
Article numbere28674
JournalPLoS One
Volume6
Issue number12
DOIs
StatePublished - Dec 8 2011

Fingerprint

Chemical stability
Chromophores
absorbance
photostability
Luminance
energy transfer
green fluorescent protein
chemical structure
Proteins
Confocal microscopy
Energy Transfer
Green Fluorescent Proteins
Confocal Microscopy
Energy transfer
Fusion reactions
proteins
cells
fluorescent proteins
protein tagging
confocal microscopy

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

An enhanced monomeric blue fluorescent protein with the high chemical stability of the chromophore. / Subach, Oksana M.; Cranfill, Paula J.; Davidson, Michael W.; Verkhusha, Vladislav.

In: PLoS One, Vol. 6, No. 12, e28674, 08.12.2011.

Research output: Contribution to journalArticle

Subach, Oksana M. ; Cranfill, Paula J. ; Davidson, Michael W. ; Verkhusha, Vladislav. / An enhanced monomeric blue fluorescent protein with the high chemical stability of the chromophore. In: PLoS One. 2011 ; Vol. 6, No. 12.
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