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Related Experiment Video

Updated: Apr 8, 2026

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Published on: April 27, 2018

Fluorescence imaging using synthetic GFP chromophores.

Christopher L Walker¹, Konstantin A Lukyanov², Ilia V Yampolsky³

¹School of Chemistry and Biochemistry, Georgia Institute of Technology, 901 Atlantic Drive, Atlanta, GA 30332-0400, United States.

Current Opinion in Chemical Biology

|June 29, 2015

Summary

Synthetic green fluorescent protein (GFP) chromophores become fluorescent when bound to host molecules. This enables their use as sensitive fluorescent dyes for detecting various biological targets and for live-cell imaging applications.

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Area of Science:

Biochemistry
Molecular Biology
Chemical Biology

Background:

Green fluorescent protein (GFP) and its analogs intrinsically form fluorescent chromophores from amino acids.
Synthetic GFP chromophore analogs are typically non-fluorescent in solution due to photoinduced isomerization.
Binding to host molecules restricts internal rotation, significantly enhancing fluorescence.

Purpose of the Study:

To investigate the fluorescence properties of synthetic GFP chromophore analogs.
To explore the potential of these analogs as fluorogenic dyes for molecular detection.
To evaluate their utility in biological imaging applications.

Main Methods:

Synthesis of GFP chromophore analogs.
Spectroscopic analysis of fluorescence properties in solution and when bound to host molecules.
Application of analogs with RNA aptamers (e.g., Spinach) for live-cell imaging.

Main Results:

Binding to host molecules induced a significant increase (up to 1000-fold) in fluorescence intensity for synthetic GFP chromophores.
Demonstrated utility of these analogs as fluorogenic dyes for detecting metal ions, proteins, and nucleic acids.
Successful application of Spinach aptamer and GFP chromophores for live-cell imaging of RNA, metabolites, and proteins.

Conclusions:

Restricting internal rotation of synthetic GFP chromophores dramatically enhances their fluorescence.
These analogs are promising as small, water-soluble, and brightly fluorescent dyes for biological sensing and imaging.
Chemically locked GFP chromophores offer a versatile platform for developing novel fluorescent probes.