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Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids

The ability to modulate protein function through minimal perturbations to amino acid structure represents an ideal mechanism to engineer optimized proteins. Due to the novel spectroscopic properties of green fluorescent protein, it has found widespread application as a reporter protein throughout th...

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Autores principales: Villa, Jordan K., Tran, Hong-Anh, Vipani, Megha, Gianturco, Stephanie, Bhasin, Konark, Russell, Brent L., Harbron, Elizabeth J., Young, Douglas D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5806519/
https://www.ncbi.nlm.nih.gov/pubmed/28714902
http://dx.doi.org/10.3390/molecules22071194
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author Villa, Jordan K.
Tran, Hong-Anh
Vipani, Megha
Gianturco, Stephanie
Bhasin, Konark
Russell, Brent L.
Harbron, Elizabeth J.
Young, Douglas D.
author_facet Villa, Jordan K.
Tran, Hong-Anh
Vipani, Megha
Gianturco, Stephanie
Bhasin, Konark
Russell, Brent L.
Harbron, Elizabeth J.
Young, Douglas D.
author_sort Villa, Jordan K.
collection PubMed
description The ability to modulate protein function through minimal perturbations to amino acid structure represents an ideal mechanism to engineer optimized proteins. Due to the novel spectroscopic properties of green fluorescent protein, it has found widespread application as a reporter protein throughout the fields of biology and chemistry. Using site-specific amino acid mutagenesis, we have incorporated various fluorotyrosine residues directly into the fluorophore of the protein, altering the fluorescence and shifting the pKa of the phenolic proton associated with the fluorophore. Relative to wild type GFP, the fluorescence spectrum of the protein is altered with each additional fluorine atom, and the mutant GFPs have the potential to be employed as pH sensors due to the altered electronic properties of the fluorine atoms.
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spelling pubmed-58065192018-02-09 Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids Villa, Jordan K. Tran, Hong-Anh Vipani, Megha Gianturco, Stephanie Bhasin, Konark Russell, Brent L. Harbron, Elizabeth J. Young, Douglas D. Molecules Communication The ability to modulate protein function through minimal perturbations to amino acid structure represents an ideal mechanism to engineer optimized proteins. Due to the novel spectroscopic properties of green fluorescent protein, it has found widespread application as a reporter protein throughout the fields of biology and chemistry. Using site-specific amino acid mutagenesis, we have incorporated various fluorotyrosine residues directly into the fluorophore of the protein, altering the fluorescence and shifting the pKa of the phenolic proton associated with the fluorophore. Relative to wild type GFP, the fluorescence spectrum of the protein is altered with each additional fluorine atom, and the mutant GFPs have the potential to be employed as pH sensors due to the altered electronic properties of the fluorine atoms. MDPI 2017-07-16 /pmc/articles/PMC5806519/ /pubmed/28714902 http://dx.doi.org/10.3390/molecules22071194 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Villa, Jordan K.
Tran, Hong-Anh
Vipani, Megha
Gianturco, Stephanie
Bhasin, Konark
Russell, Brent L.
Harbron, Elizabeth J.
Young, Douglas D.
Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids
title Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids
title_full Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids
title_fullStr Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids
title_full_unstemmed Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids
title_short Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids
title_sort fluorescence modulation of green fluorescent protein using fluorinated unnatural amino acids
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5806519/
https://www.ncbi.nlm.nih.gov/pubmed/28714902
http://dx.doi.org/10.3390/molecules22071194
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