Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2019 Aug 12;9(1):11640.
doi: 10.1038/s41598-019-47660-0.

Role of Hydrogen Bonding in Green Fluorescent Protein-like Chromophore Emission

Li Yang  1   2 , Shifeng Nian  2 , Guozhen Zhang  2 , Edward Sharman  3 , Hui Miao  2 , Xuepeng Zhang  4 , Xiaofeng Chen  5 , Yi Luo  2 , Jun Jiang  6
Affiliations

Role of Hydrogen Bonding in Green Fluorescent Protein-like Chromophore Emission

Li Yang et al. Sci Rep. .

Abstract

The fluorescence emission from green fluorescent protein (GFP) is known to be heavily influenced by hydrogen bonding between the core fluorophore and the surrounding side chains or water molecules. Yet how to utilize this feature for modulating the fluorescence of GFP chromophore or GFP-like fluorophore still remains elusive. Here we present theoretical calculations to predict how hydrogen bonding could influence the excited states of the GFP-like fluorophores. These studies provide both a new perspective for understanding the photophysical properties of GFP as well as a solid basis for the rational design of GFP-based fluorophores.

PubMed Disclaimer

Conflict of interest statement

The authors declare no competing interests.

Figures

Figure 1
Figure 1
Photo absorption and emission process of the modified GFP chromophore. (a) Chemical structures of the GFP chromophore and (b) the C=C double bond-modified GFP chromophore. (c) The computed absorption spectrum together with the lowest photo-emission peak (d) of the modified chromophore. The calculated transitions between frontier orbitals for the photo absorption (e) and emission (f) of the modified chromophore in aqueous solution. The blue, gray, white and red balls represent N, C, H and O atoms, respectively. f stands for oscillator strength.
Figure 2
Figure 2
Fluorescent redshift of the modified chromophore with hydrogen bonding network consisting of hydrogen and water. Molecular configurations of the modified chromophore with hydrogen in the (a) absence and (b) presence of water molecules, added hydrogen atom are highlighted in blue. (c) The computed electronic energy levels and photo-emission transitions of the two excited-state structures and the original modified chromophore. The orbital transitions for the calculated photo-emission peak of the chromophore with hydrogen in the (d) absence and (e) presence of water molecules in aqueous solution. The green dotted bonds denote hydrogen bonding.
Figure 3
Figure 3
Optical properties of the cationic protonated modified chromophore and that with water molecule. Molecular configurations of the monocationic protonated modified structure in the (a) absence and (b) presence of water molecule, added protons are highlighted in blue. (c) The computed electronic energy levels and photo-emission transitions of these two excited-state structures and the original modified chromophore. The calculated transitions between frontier orbitals for the photo-emission peak of the protonated cation in the (d) absence and (e) presence of a water molecule.
Figure 4
Figure 4
Relationship between the emission wavelengths and relevant bond lengths. The calculated emission maximum as a function of selected bond lengths for the modified chromophore with hydrogen in the (a) absence and (b) presence of water molecules in aqueous solution.
See this image and copyright information in PMC

References

    1. Chalfie M, Tu Y, Euskirchen G, Ward WW, Prasher DC. Green fluorescent protein as a marker for gene expression. Science. 1994;263:802. doi: 10.1126/science.8303295. - DOI - PubMed
    1. Miyawaki A, et al. Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin. Nature. 1997;388:882–887. doi: 10.1038/42264. - DOI - PubMed
    1. Patterson GH, Lippincott-Schwartz J. A Photoactivatable GFP for Selective Photolabeling of Proteins and Cells. Science. 2002;297:1873. doi: 10.1126/science.1074952. - DOI - PubMed
    1. Sattarzadeh A, Saberianfar R, Zipfel WR, Menassa R, Hanson MR. Green to red photoconversion of GFP for protein tracking in vivo. Sci. Rep. 2015;5:11771–11771. doi: 10.1038/srep11771. - DOI - PMC - PubMed
    1. Giepmans BNG, Adams SR, Ellisman MH, Tsien RY. The Fluorescent Toolbox for Assessing Protein Location and Function. Science. 2006;312:217. doi: 10.1126/science.1124618. - DOI - PubMed

Publication types