Design principles of green fluorescent proteins - Why are they green and fluorescent?
Abstract/Contents
- Abstract
- Since its discovery in jellyfish around the early 1960s, green fluorescent protein (GFP) and its derivatives have become the most widely used in vivo imaging tools for biological studies. Mutants of GFPs have been engineered to exhibit a palette of colors, wide range of fluorescence quantum yields (FQYs), and photoswitching characteristics, enabling advanced applications such as optogenetics and super-resolution microscopy. However, tailoring GFPs for specific purposes heavily relies on extensive screening and/or directed evolution as it is unclear how the protein environment modulates the photophysics of the chromophore in quantitative terms. Thanks to the semisynthetic split GFP method and the recently popularized technique for incorporating noncanonical amino acids (i.e., amber suppression), we created conventionally inaccessible GFP mutants with systematically altered electrostatic properties and/or steric bulk of the chromophore and the surrounding protein environment. Correlations between various photophysical properties of these mutants led to transparent models describing how the protein-chromophore interactions affect the chromophore's color and FQY. The models provide quantitative predictions for designing GFPs with desired phenotypes and infer physically imposed limitations. In addition, our finding of how electrostatics biases photoisomerization pathways in GFPs could shed light on the general phenomenon of bond-specific photoisomerization, a defining characteristic of the retinal chromophore within rhodopsins central to vision and important to the field of photoactive molecular devices.
Description
| Type of resource | text |
|---|---|
| Form | electronic resource; remote; computer; online resource |
| Extent | 1 online resource. |
| Place | California |
| Place | [Stanford, California] |
| Publisher | [Stanford University] |
| Copyright date | 2020; ©2020 |
| Publication date | 2020; 2020 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Author | Lin, Chi-Yun |
|---|---|
| Degree supervisor | Boxer, Steven G. (Steven George), 1947- |
| Thesis advisor | Boxer, Steven G. (Steven George), 1947- |
| Thesis advisor | Cui, Bianxiao |
| Thesis advisor | Martinez, Todd J. (Todd Joseph), 1968- |
| Degree committee member | Cui, Bianxiao |
| Degree committee member | Martinez, Todd J. (Todd Joseph), 1968- |
| Associated with | Stanford University, Department of Chemistry. |
Subjects
| Genre | Theses |
|---|---|
| Genre | Text |
Bibliographic information
| Statement of responsibility | Chi-Yun Lin. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Thesis Ph.D. Stanford University 2020. |
| Location | electronic resource |
Access conditions
- Copyright
- © 2020 by Chi-Yun Lin
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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