Molecular Basis of Spectral Diversity in Near-Infrared Phytochrome-Based Fluorescent Proteins

Daria Shcherbakova, Mikhail Baloban, Sergei Pletnev, Vladimir N. Malashkevich, Hui Xiao, Zbigniew Dauter, Vladislav Verkhusha

Research output: Contribution to journalArticle

27 Citations (Scopus)

Abstract

Summary Near-infrared fluorescent proteins (NIR FPs) engineered from bacterial phytochromes (BphPs) are the probes of choice for deep-tissue imaging. Detection of several processes requires spectrally distinct NIR FPs. We developed an NIR FP, BphP1-FP, which has the most blue-shifted spectra and the highest fluorescence quantum yield among BphP-derived FPs. We found that these properties result from the binding of the biliverdin chromophore to a cysteine residue in the GAF domain, unlike natural BphPs and other BphP-based FPs. To elucidate the molecular basis of the spectral shift, we applied biochemical, structural and mass spectrometry analyses and revealed the formation of unique chromophore species. Mutagenesis of NIR FPs of different origins indicated that the mechanism of the spectral shift is general and can be used to design multicolor NIR FPs from other BphPs. We applied pairs of spectrally distinct point cysteine mutants to multicolor cell labeling and demonstrated that they perform well in model deep-tissue imaging.

Original languageEnglish (US)
Pages (from-to)1540-1551
Number of pages12
JournalChemistry and Biology
Volume22
Issue number11
DOIs
StatePublished - Nov 19 2015

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Phytochrome
Infrared radiation
Cysteine
Chromophores
Biliverdine
Proteins
Bacterial Proteins
Tissue
Imaging techniques
Mutagenesis
Mass Spectrometry
Quantum yield
Fluorescence
Labeling
Mass spectrometry

ASJC Scopus subject areas

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

Cite this

Molecular Basis of Spectral Diversity in Near-Infrared Phytochrome-Based Fluorescent Proteins. / Shcherbakova, Daria; Baloban, Mikhail; Pletnev, Sergei; Malashkevich, Vladimir N.; Xiao, Hui; Dauter, Zbigniew; Verkhusha, Vladislav.

In: Chemistry and Biology, Vol. 22, No. 11, 19.11.2015, p. 1540-1551.

Research output: Contribution to journalArticle

Shcherbakova, Daria ; Baloban, Mikhail ; Pletnev, Sergei ; Malashkevich, Vladimir N. ; Xiao, Hui ; Dauter, Zbigniew ; Verkhusha, Vladislav. / Molecular Basis of Spectral Diversity in Near-Infrared Phytochrome-Based Fluorescent Proteins. In: Chemistry and Biology. 2015 ; Vol. 22, No. 11. pp. 1540-1551.
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