Open access
Date
2012-10-31Type
- Journal Article
ETH Bibliography
yes
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Abstract
Green fluorescent protein (GFP) mutants have become the most widely used fluorescence markers in the life sciences, and although they are becoming increasingly popular as mechanical force or strain probes, there is little direct information on how their fluorescence changes when mechanically stretched. Here we derive high-resolution structural models of the mechanical intermediate states of stretched GFP using steered molecular dynamics (SMD) simulations. These structures were used to produce mutants of EGFP and EYFP that mimic GFP's different mechanical intermediates. A spectroscopic analysis revealed that a population of EGFP molecules with a missing N-terminal α-helix was significantly dimmed, while the fluorescence lifetime characteristic of the anionic chromophore state remained unaffected. This suggests a mechanism how N-terminal deletions can switch the protonation state of the chromophore, and how the fluorescence of GFP molecules in response to mechanical disturbance might be turned off. Show more
Permanent link
https://doi.org/10.3929/ethz-b-000058799Publication status
publishedExternal links
Journal / series
PLoS ONEVolume
Pages / Article No.
Publisher
PLOSOrganisational unit
03640 - Vogel, Viola / Vogel, Viola
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ETH Bibliography
yes
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