TY - JOUR
T1 - Smallest near-infrared fluorescent protein evolved from cyanobacteriochrome as versatile tag for spectral multiplexing
AU - Oliinyk, Olena S.
AU - Shemetov, Anton A.
AU - Pletnev, Sergei
AU - Shcherbakova, Daria M.
AU - Verkhusha, Vladislav V.
N1 - Funding Information:
We thank K. Aoki (Okazaki Institute for Integrative Bioscience, Japan) and M. Matsuda (Kyoto University, Japan) for the AKAR3EV and JNKAR1EV plasmids, A. Kaberniuk from our laboratory for the help with animal experiments, N. Peitsaro and N. Aarnio from Flow Cytometry Core Facility and staff of Neuronal Cell Culture Unit of the University of Helsinki for the technical assistance. This work was supported by grants GM122567 and NS103573 from the US National Institutes of Health (NIH), and ERC-2013-ADG-340233 from the EU FP7 program. This work was also funded in part with federal funds from the Frederick National Laboratory for Cancer Research, NIH contract HHSN261200800001E, and the Intramural Research Program of the NIH, Frederick National Laboratory, Center for Cancer Research. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the US Government.
Funding Information:
1Medicum, Faculty of Medicine, University of Helsinki, 00290 Helsinki, Finland. 2Department of Anatomy and Structural Biology, and Gruss-Lipper Biophotonics Center, Albert Einstein College of Medicine, Bronx, NY 10461, USA. 3Basic Science Program, Macromolecular Crystallography Laboratory, Frederick National Laboratory for Cancer Research sponsored by the National Cancer Institute, Frederick, MD 21702, USA. Correspondence and requests for materials should be addressed to V.V.V. (email: vladislav.verkhusha@einstein.yu.edu)
Publisher Copyright:
© 2019, The Author(s).
PY - 2019/12/1
Y1 - 2019/12/1
N2 - 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.
AB - 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.
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U2 - 10.1038/s41467-018-08050-8
DO - 10.1038/s41467-018-08050-8
M3 - Article
C2 - 30655515
AN - SCOPUS:85060171389
SN - 2041-1723
VL - 10
JO - Nature Communications
JF - Nature Communications
IS - 1
M1 - 279
ER -