5DTX

Crystal structure of rsEGFP2 in the fluorescent on-state

Summary for 5DTX

• Entry DOI: 10.2210/pdb5dtx/pdb
• Descriptor: Green fluorescent protein (2 entities in total)
• Functional Keywords: fluorescent protein, gfp, reversibly switchable, cis chromophore
• Biological source: Aequorea victoria (Jellyfish)
• Total number of polymer chains: 1
• Total formula weight: 28099.69

Authors

Adam, V.,Martins, A. (deposition date: 2015-09-18, release date: 2016-01-20, Last modification date: 2024-01-10)

Primary citation

El Khatib, M.,Martins, A.,Bourgeois, D.,Colletier, J.P.,Adam, V.
Rational design of ultrastable and reversibly photoswitchable fluorescent proteins for super-resolution imaging of the bacterial periplasm.
Sci Rep, 6:18459-18459, 2016

PubMed Abstract: Phototransformable fluorescent proteins are central to several nanoscopy approaches. As yet however, there is no available variant allowing super-resolution imaging in cell compartments that maintain oxidative conditions. Here, we report the rational design of two reversibly switchable fluorescent proteins able to fold and photoswitch in the bacterial periplasm, rsFolder and rsFolder2. rsFolder was designed by hybridisation of Superfolder-GFP with rsEGFP2, and inherited the fast folding properties of the former together with the rapid switching of the latter, but at the cost of a reduced switching contrast. Structural characterisation of the switching mechanisms of rsFolder and rsEGFP2 revealed different scenarios for chromophore cis-trans isomerisation and allowed designing rsFolder2, a variant of rsFolder that exhibits improved switching contrast and is amenable to RESOLFT nanoscopy. The rsFolders can be efficiently expressed in the E. coli periplasm, opening the door to the nanoscale investigation of proteins localised in hitherto non-observable cellular compartments.

PubMed: 26732634
DOI: 10.1038/srep18459

Experimental method

X-RAY DIFFRACTION (1.45 Å)