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...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2017-07-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/22/7/1194 |
| _version_ | 1818423287673257984 |
|---|---|
| author | Jordan K. Villa Hong-Anh Tran Megha Vipani Stephanie Gianturco Konark Bhasin Brent L. Russell Elizabeth J. Harbron Douglas D. Young |
| author_facet | Jordan K. Villa Hong-Anh Tran Megha Vipani Stephanie Gianturco Konark Bhasin Brent L. Russell Elizabeth J. Harbron Douglas D. Young |
| author_sort | Jordan K. Villa |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-12-14T13:39:45Z |
| format | Article |
| id | doaj.art-0e779d6aade64233a44ce805c4e2bdf6 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-12-14T13:39:45Z |
| publishDate | 2017-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-0e779d6aade64233a44ce805c4e2bdf62022-12-21T22:59:29ZengMDPI AGMolecules1420-30492017-07-01227119410.3390/molecules22071194molecules22071194Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino AcidsJordan K. Villa0Hong-Anh Tran1Megha Vipani2Stephanie Gianturco3Konark Bhasin4Brent L. Russell5Elizabeth J. Harbron6Douglas D. Young7Department of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USADepartment of Chemistry, The College of William & Mary, Williamsburg, VA 231871, USAThe 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.https://www.mdpi.com/1420-3049/22/7/1194unnatural amino acidsgreen fluorescent proteinbiosensorsfluorotyrosine |
| spellingShingle | Jordan K. Villa Hong-Anh Tran Megha Vipani Stephanie Gianturco Konark Bhasin Brent L. Russell Elizabeth J. Harbron Douglas D. Young Fluorescence Modulation of Green Fluorescent Protein Using Fluorinated Unnatural Amino Acids Molecules unnatural amino acids green fluorescent protein biosensors fluorotyrosine |
| 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 | unnatural amino acids green fluorescent protein biosensors fluorotyrosine |
| url | https://www.mdpi.com/1420-3049/22/7/1194 |
| work_keys_str_mv | AT jordankvilla fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT honganhtran fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT meghavipani fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT stephaniegianturco fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT konarkbhasin fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT brentlrussell fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT elizabethjharbron fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids AT douglasdyoung fluorescencemodulationofgreenfluorescentproteinusingfluorinatedunnaturalaminoacids |