Mécanismes de photo-commutation réversible des protéines fluorescentes

Abstract : The property to be reversible switchable of some homologues fluorescents protein ofGFP open a large field for possible applications: such as, high-density data bio-storage andsuper-resolution microscopy. Between these proteins, we find several variants of GFP, such asyellow fluorescent protein, YFP, and fluorescents protein from marine Anthozoary species, asDronpa or Padron. Several structural studies suggest that these fluorescent proteins switch viaisomerization coupled with the protonation of the chromophore. However, thesynchronization between these processes, the detail about the photo-switching mechanism,and the role of conformational dynamics remains unclear. In combination of the kineticcrystallography and the optic spectroscopy in cristallo at low temperature, we have comparedthe YFP behavior, Dronpa and IrisFP, and we have studied in detail the photo-physicmechanism of Padron switching. In contrast to Dronpa and IrisFP, the YFP photoswitching ismore efficient at low temperature than at room temperature. Our results suggest that theYFPswitching is not associated to large structural rearrangements, but mostly a photo-inducedprotonation of the chromophore without isomerization. On the contrary, the studies done withPadron allowed us to show, in this case, the chromophore isomerization can be producedindependently of the protonation, at cryo-temperatures. Moreover, two intermediate stateswere revealed in the photo-pathway. Padron fluorescent protein allows to advance the firstgenetically inserted dye, being photo-switchable at cryogenic temperature
Complete list of metadatas

https://tel.archives-ouvertes.fr/tel-00858895
Contributor : Abes Star <>
Submitted on : Friday, September 6, 2013 - 11:22:49 AM
Last modification on : Friday, April 5, 2019 - 1:01:47 AM
Long-term archiving on : Saturday, December 7, 2013 - 4:17:48 AM

File

24866_FARO_2012_archivage.pdf
Version validated by the jury (STAR)

Identifiers

  • HAL Id : tel-00858895, version 1

Collections

Citation

Aline Regis Faro. Mécanismes de photo-commutation réversible des protéines fluorescentes. Autre [cond-mat.other]. Université de Grenoble, 2012. Français. ⟨NNT : 2012GRENY056⟩. ⟨tel-00858895⟩

Share

Metrics

Record views

674

Files downloads

902