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Updated: Jun 25, 2026
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Time-Resolved Fluorescence Anisotropy from Single Molecules for Characterizing Local Flexibility in Biomolecules
Published on: April 25, 2025
Structure, dynamics and optical properties of fluorescent proteins: perspectives for marker development
G Ulrich Nienhaus1, Jörg Wiedenmann
1Institute of Biophysics, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany. uli@uiuc.edu
Summary
Green fluorescent proteins (GFPs) and similar markers are vital for life science research. Understanding their structure and dynamics helps engineer advanced fluorescent proteins, especially red fluorescent variants, for better cellular imaging.
Area of Science:
- Biochemistry
- Molecular Biology
- Biophysics
Background:
- Genetically encoded fluorescence markers, like Green Fluorescent Proteins (GFPs), are essential tools in life sciences, derived from marine organisms.
- Ongoing research seeks novel GFP-like proteins with improved properties for advanced cellular imaging applications.
- Variations in fluorophore structure and the surrounding protein environment significantly influence optical characteristics.
Purpose of the Study:
- To review the structural diversity of GFP-like proteins.
- To discuss the relationship between protein structure, dynamics, and optical properties.
- To highlight advancements in red fluorescent proteins and photoactivatable markers.
Main Methods:
- Structural analysis of GFP-like proteins.
- Investigation of fluorophore covalent structure and stereochemistry.
- Examination of protein dynamics and solvation shell effects on optical properties.
Main Results:
- Significant structural variations exist within GFP-like proteins, impacting their optical properties.
- The fluorophore's microenvironment, including its solvation shell, modulates absorption/emission wavelengths and brightness.
- Recent progress includes the development of photoactivatable fluorescence markers with tunable properties.
Conclusions:
- A comprehensive understanding of GFP-like protein structure and dynamics is crucial for rational design.
- Engineering advanced fluorescence markers relies on detailed knowledge of structure-property relationships.
- Further development, particularly for red fluorescent proteins, promises enhanced capabilities in cellular imaging.
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