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Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing

Author

Listed:
  • Olena S. Oliinyk

    (University of Helsinki)

  • Anton A. Shemetov

    (Albert Einstein College of Medicine)

  • Sergei Pletnev

    (Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute)

  • Daria M. Shcherbakova

    (Albert Einstein College of Medicine)

  • Vladislav V. Verkhusha

    (University of Helsinki
    Albert Einstein College of Medicine)

Abstract

From a single domain of cyanobacteriochrome (CBCR) we developed a near-infrared (NIR) fluorescent protein (FP), termed miRFP670nano, with excitation at 645 nm and emission at 670 nm. This is the first CBCR-derived NIR FP evolved to efficiently bind endogenous biliverdin chromophore and brightly fluoresce in mammalian cells. miRFP670nano is a monomer with molecular weight of 17 kDa that is 2-fold smaller than bacterial phytochrome (BphP)-based NIR FPs and 1.6-fold smaller than GFP-like FPs. Crystal structure of the CBCR-based NIR FP with biliverdin reveals a molecular basis of its spectral and biochemical properties. Unlike BphP-derived NIR FPs, miRFP670nano is highly stable to denaturation and degradation and can be used as an internal protein tag. miRFP670nano is an effective FRET donor for red-shifted NIR FPs, enabling engineering NIR FRET biosensors spectrally compatible with GFP-like FPs and blue–green optogenetic tools. miRFP670nano unlocks a new source of diverse CBCR templates for NIR FPs.

Suggested Citation

  • Olena S. Oliinyk & Anton A. Shemetov & Sergei Pletnev & Daria M. Shcherbakova & Vladislav V. Verkhusha, 2019. "Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing," Nature Communications, Nature, vol. 10(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-018-08050-8
    DOI: 10.1038/s41467-018-08050-8
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    Cited by:

    1. Natalia V. Barykina & Erin M. Carey & Olena S. Oliinyk & Axel Nimmerjahn & Vladislav V. Verkhusha, 2024. "Destabilized near-infrared fluorescent nanobodies enable background-free targeting of GFP-based biosensors for imaging and manipulation," Nature Communications, Nature, vol. 15(1), pages 1-16, December.

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