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

Olena S. Oliinyk, Anton A. Shemetov, Sergei Pletnev, Daria M. Shcherbakova and Vladislav V. Verkhusha ()
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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

Nature Communications, 2019, vol. 10, issue 1, 1-13

Abstract: 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.

Date: 2019
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DOI: 10.1038/s41467-018-08050-8

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