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

Show full item record



Permalink

http://hdl.handle.net/10138/197742

Citation

Baloban , M , Shcherbakova , D M , Pletnev , S , Pletnev , V Z , Lagarias , J C & Verkhusha , V 2017 , ' Designing brighter near-infrared fluorescent proteins : insights from structural and biochemical studies ' , Chemical Science , vol. 8 , no. 6 , pp. 4546-4557 . https://doi.org/10.1039/c7sc00855d

Title: Designing brighter near-infrared fluorescent proteins : insights from structural and biochemical studies
Author: Baloban, Mikhail; Shcherbakova, Daria M.; Pletnev, Sergei; Pletnev, Vladimir Z.; Lagarias, J. Clark; Verkhusha, Vladislav
Contributor: University of Helsinki, Medicum
Date: 2017-06
Language: eng
Number of pages: 12
Belongs to series: Chemical Science
ISSN: 2041-6520
URI: http://hdl.handle.net/10138/197742
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.
Subject: CHROMOPHORE-BINDING DOMAIN
AGROBACTERIUM-PHYTOCHROME AGP1
EXCITED-STATE DYNAMICS
IN-VIVO
BACTERIAL PHYTOCHROMES
DEINOCOCCUS-RADIODURANS
BILIVERDIN CHROMOPHORE
MOLECULAR REPLACEMENT
CRYSTAL-STRUCTURE
QUANTUM YIELD
3111 Biomedicine
1182 Biochemistry, cell and molecular biology
116 Chemical sciences
Rights:


Files in this item

Total number of downloads: Loading...

Files Size Format View
c7sc00855d.pdf 1.430Mb PDF View/Open

This item appears in the following Collection(s)

Show full item record