Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids

被引:15
作者
Villa, Jordan K. [1 ]
Hong-Anh Tran [1 ]
Vipani, Megha [1 ]
Gianturco, Stephanie [1 ]
Bhasin, Konark [1 ]
Russell, Brent L. [1 ]
Harbron, Elizabeth J. [1 ]
Young, Douglas D. [1 ]
机构
[1] Coll William & Mary, Dept Chem, Williamsburg, VA 23187 USA
关键词
unnatural amino acids; green fluorescent protein; biosensors; fluorotyrosine; SITE-SPECIFIC INCORPORATION; TRANSFER-RNA SYNTHETASE; ESCHERICHIA-COLI; GENETIC-CODE; RIBONUCLEOTIDE REDUCTASE; GFP; FLUOROTYROSINES; LIGATION;
D O I
10.3390/molecules22071194
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
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页数:7
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