Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome

Kiryl D. Piatkevich, Fedor V. Subach, Vladislav Verkhusha

Research output: Contribution to journalArticle

64 Citations (Scopus)

Abstract

The ability to modulate the fluorescence of optical probes can be used to enhance signal-to-noise ratios for imaging within highly autofluorescent environments, such as intact tissues and living organisms. Here, we report two bacteriophytochrome-based photoactivatable near-infrared fluorescent proteins, named PAiRFP1 and PAiRFP2. PAiRFPs utilize haem-derived biliverdin, ubiquitous in mammalian tissues, as the chromophore. Initially weakly fluorescent PAiRFPs undergo photoconversion into a highly fluorescent state with excitation/emission at 690/717 nm following a brief irradiation with far-red light. After photoactivation, PAiRFPs slowly revert back to initial state, enabling multiple photoactivation-relaxation cycles. Low-temperature optical spectroscopy reveals several intermediates involved in PAiRFP photocycles, which all differ from that of the bacteriophytochrome precursor. PAiRFPs can be photoactivated in a spatially selective manner in mouse tissues, and optical modulation of their fluorescence allows for substantial contrast enhancement, making PAiRFPs advantageous over permanently fluorescent probes for in vivo imaging conditions of high autofluorescence and low signal levels.

Original languageEnglish (US)
Article number2153
JournalNature Communications
Volume4
DOIs
StatePublished - 2013

Fingerprint

phytochrome
Phytochrome
Tissue
Infrared radiation
proteins
Light
Fluorescence
Biliverdine
Imaging techniques
fluorescence
Proteins
probes
Light modulation
light modulation
Signal-To-Noise Ratio
Chromophores
Heme
Fluorescent Dyes
organisms
chromophores

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Chemistry(all)
  • Physics and Astronomy(all)

Cite this

Far-red light photoactivatable near-infrared fluorescent proteins engineered from a bacterial phytochrome. / Piatkevich, Kiryl D.; Subach, Fedor V.; Verkhusha, Vladislav.

In: Nature Communications, Vol. 4, 2153, 2013.

Research output: Contribution to journalArticle

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