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Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids.

ABSTRACT: 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.

SUBMITTER: Villa JK

PROVIDER: S-EPMC5806519 | biostudies-literature | 2017 Jul

REPOSITORIES: biostudies-literature

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Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids.

Villa Jordan K JK Tran Hong-Anh HA Vipani Megha M Gianturco Stephanie S Bhasin Konark K Russell Brent L BL Harbron Elizabeth J EJ Young Douglas D DD

Molecules (Basel, Switzerland) 20170716 7

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 fluoresce ...[more]

PMID: 28714902

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