Crystal Structure of Bright Fluorescent Protein BrUSLEE with Subnanosecond Fluorescence Lifetime; Electric and Dynamic Properties

The rapid development of new microscopy techniques for cell biology has exposed the need for genetically encoded fluorescent tags with special properties. Fluorescent biomarkers of the same color and spectral range and different fluorescent lifetimes (FLs) became useful for fluorescent lifetime imag...

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Bibliographic Details
Published in:International journal of molecular sciences 2023-03, Vol.24 (7), p.6403
Main Authors: Goryacheva, Ekaterina, Efremov, Roman, Krylov, Nikolai, Artemyev, Igor, Bogdanov, Alexey, Mamontova, Anastasia, Pletnev, Sergei, Pletneva, Nadya, Pletnev, Vladimir
Format: Article
Language:English
Subjects:
Aqueous solutions
Binding sites
Cells
Chromophores
Coloring Agents
computer modeling
Crystal structure
Crystals
Dynamic structural analysis
Electric properties
Emitters
Fluorescence
fluorescence lifetime
fluorescent protein
Genetic code
Green fluorescent protein
Green Fluorescent Proteins - metabolism
Hydrogen bonding
Lifetime
Microenvironments
Microscopy
Microscopy, Fluorescence - methods
Molecular conformation
Molecular dynamics
Mutation
Protein structure
Proteins
Service life assessment
Spectrum analysis
Structure
X-ray structure
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Summary:The rapid development of new microscopy techniques for cell biology has exposed the need for genetically encoded fluorescent tags with special properties. Fluorescent biomarkers of the same color and spectral range and different fluorescent lifetimes (FLs) became useful for fluorescent lifetime image microscopy (FLIM). One such tag, the green fluorescent protein BrUSLEE (Bright Ultimately Short Lifetime Enhanced Emitter), having an extremely short subnanosecond component of fluorescence lifetime (FL~0.66 ns) and exceptional fluorescence brightness, was designed for FLIM experiments. Here, we present the X-ray structure and discuss the structure-functional relations of BrUSLEE. Its development from the EGFP (enhanced green fluorescent proteins) precursor (FL~2.83 ns) resulted in a change of the chromophore microenvironment due to a significant alteration in the side chain conformations. To get further insight into molecular details explaining the observed differences in the photophysical properties of these proteins, we studied their structural, dynamic, and electric properties by all-atom molecular-dynamics simulations in an aqueous solution. It has been shown that compared to BrUSLEE, the mobility of the chromophore in the EGFP is noticeably limited by nonbonded interactions (mainly H-bonds) with the neighboring residues.
ISSN:1422-0067
1661-6596
1422-0067
DOI:10.3390/ijms24076403