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Issue 6, 2017
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Designing brighter near-infrared fluorescent proteins: insights from structural and biochemical studies

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Abstract

Brighter near-infrared (NIR) fluorescent proteins (FPs) are required for multicolor microscopy and deep-tissue imaging. Here, we present structural and biochemical analyses of three monomeric, spectrally distinct phytochrome-based NIR FPs, termed miRFPs. The miRFPs are closely related and differ by only a few amino acids, which define their molecular brightness, brightness in mammalian cells, and spectral properties. We have identified the residues responsible for the spectral red-shift, revealed a new chromophore bound simultaneously to two cysteine residues in the PAS and GAF domains in blue-shifted NIR FPs, and uncovered the importance of amino acid residues in the N-terminus of NIR FPs for their molecular and cellular brightness. The novel chromophore covalently links the N-terminus of NIR FPs with their C-terminal GAF domain, forming a topologically closed knot in the structure, and also contributes to the increased brightness. Based on our studies, we suggest a strategy to develop spectrally distinct NIR FPs with enhanced brightness.

Graphical abstract: Designing brighter near-infrared fluorescent proteins: insights from structural and biochemical studies

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Publication details

The article was received on 23 Feb 2017, accepted on 11 Apr 2017 and first published on 04 May 2017


Article type: Edge Article
DOI: 10.1039/C7SC00855D
Citation: Chem. Sci., 2017,8, 4546-4557
  • Open access: Creative Commons BY-NC license
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    Designing brighter near-infrared fluorescent proteins: insights from structural and biochemical studies

    M. Baloban, D. M. Shcherbakova, S. Pletnev, V. Z. Pletnev, J. C. Lagarias and V. V. Verkhusha, Chem. Sci., 2017, 8, 4546
    DOI: 10.1039/C7SC00855D

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