Designing brighter near-infrared fluorescent proteins: Insights from structural and biochemical studies

Mikhail Baloban, Daria Shcherbakova, Sergei Pletnev, Vladimir Z. Pletnev, J. Clark Lagarias, Vladislav Verkhusha

Research output: Contribution to journalArticle

15 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)4546-4557
Number of pages12
JournalChemical Science
Volume8
Issue number6
DOIs
StatePublished - 2017

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Luminance
Infrared radiation
Proteins
Chromophores
Phytochrome
Amino Acids
Cysteine
Microscopic examination
Cells
Tissue
Imaging techniques

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Designing brighter near-infrared fluorescent proteins : Insights from structural and biochemical studies. / Baloban, Mikhail; Shcherbakova, Daria; Pletnev, Sergei; Pletnev, Vladimir Z.; Lagarias, J. Clark; Verkhusha, Vladislav.

In: Chemical Science, Vol. 8, No. 6, 2017, p. 4546-4557.

Research output: Contribution to journalArticle

Baloban, Mikhail ; Shcherbakova, Daria ; Pletnev, Sergei ; Pletnev, Vladimir Z. ; Lagarias, J. Clark ; Verkhusha, Vladislav. / Designing brighter near-infrared fluorescent proteins : Insights from structural and biochemical studies. In: Chemical Science. 2017 ; Vol. 8, No. 6. pp. 4546-4557.
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