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 organization: | Medicum Department of Biochemistry and Developmental Biology |
Date: | 2017-06 |
Language: | eng |
Number of pages: | 12 |
Belongs to series: | Chemical Science |
ISSN: | 2041-6520 |
DOI: | https://doi.org/10.1039/c7sc00855d |
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 |
Peer reviewed: | Yes |
Rights: | cc_by_nc |
Usage restriction: | openAccess |
Self-archived version: | publishedVersion |
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