Light activated reassembly of split green fluorescent protein (GFP)
Abstract/Contents
- Abstract
- Green fluorescent protein (GFP) is an incredibly useful tool to image protein expression, location, and interactions because the chromophore forms from within the protein without need for additional enzymes or external cofactors. Split GFPs are fragments of GFP that often require fused interacting proteins in order to interact, fold, and form the chromophore and are therefore used to image protein-protein interactions specifically. In this thesis a split GFP that does not require fused interacting proteins is used to make semi-synthetic proteins. The reassembly of this particular split GFP is light mediated, and the underlying mechanism and kinetics of this reassembly process are discussed in detail. Truncated Green Fluorescent Protein with the 11th [beta]-strand removed is potentially interesting for bioconjugation, imaging, and the preparation of semi-synthetic proteins with novel spectroscopic or functional properties. Surprisingly, the truncated GFP generated by removing the 11th strand, once refolded, does not reassemble with a synthetic peptide corresponding to strand 11, but does reassemble following light activation. The chromophore in this refolded truncated GFP is found to be in the trans configuration. Upon exposure to light a photostationary state is formed between the trans and cis conformations of the chromophore, and only truncated GFP with the cis configuration of the chromophore binds the peptide. As described above, GFPs and split GFPs are typically used to observe protein-protein interactions, but this unique light-driven reassembly of split GFP is potentially useful for controlling protein-protein interactions.
Description
| Type of resource | text |
|---|---|
| Form | electronic; electronic resource; remote |
| Extent | 1 online resource. |
| Publication date | 2011 |
| Issuance | monographic |
| Language | English |
Creators/Contributors
| Associated with | Kent, Kevin Patrick |
|---|---|
| Associated with | Stanford University, Department of Chemistry |
| Primary advisor | Boxer, Steven G. (Steven George), 1947- |
| Thesis advisor | Boxer, Steven G. (Steven George), 1947- |
| Thesis advisor | Chidsey, Christopher E. D. (Christopher Elisha Dunn) |
| Thesis advisor | Rao, Jianghong |
| Advisor | Chidsey, Christopher E. D. (Christopher Elisha Dunn) |
| Advisor | Rao, Jianghong |
Subjects
| Genre | Theses |
|---|
Bibliographic information
| Statement of responsibility | Kevin Patrick Kent. |
|---|---|
| Note | Submitted to the Department of Chemistry. |
| Thesis | Ph.D. Stanford University 2011 |
| Location | electronic resource |
Access conditions
- Copyright
- © 2011 by Kevin Patrick Kent
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported license (CC BY-NC).
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