5DTX: Crystal Structure Of Rsegfp2 In The Fluorescent On-state

Citation:
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.
PDB ID: 5DTXDownload
MMDB ID: 135754
PDB Deposition Date: 2015/9/18
Updated in MMDB: 2016/01
Experimental Method:
x-ray diffraction
Resolution: 1.45  Å
Source Organism:
Similar Structures:
Biological Unit for 5DTX: monomeric; determined by author and by software (PISA)
Molecular Components in 5DTX
Label Count Molecule
Protein (1 molecule)
1
Green Fluorescent Protein
Molecule annotation
* Click molecule labels to explore molecular sequence information.

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