3WCK: Crystal Structure Of Monomeric Photosensitizing Fluorescent Protein, Supernova

Citation:
Abstract
Chromophore-assisted light inactivation (CALI) is a powerful technique for acute perturbation of biomolecules in a spatio-temporally defined manner in living specimen with reactive oxygen species (ROS). Whereas a chemical photosensitizer including fluorescein must be added to specimens exogenously and cannot be restricted to particular cells or sub-cellular compartments, a genetically-encoded photosensitizer, KillerRed, can be controlled in its expression by tissue specific promoters or subcellular localization tags. Despite of this superiority, KillerRed hasn't yet become a versatile tool because its dimerization tendency prevents fusion with proteins of interest. Here, we report the development of monomeric variant of KillerRed (SuperNova) by direct evolution using random mutagenesis. In contrast to KillerRed, SuperNova in fusion with target proteins shows proper localization. Furthermore, unlike KillerRed, SuperNova expression alone doesn't perturb mitotic cell division. Supernova retains the ability to generate ROS, and hence promote CALI-based functional analysis of target proteins overcoming the major drawbacks of KillerRed.
PDB ID: 3WCKDownload
MMDB ID: 114206
PDB Deposition Date: 2013/5/27
Updated in MMDB: 2013/10
Experimental Method:
x-ray diffraction
Resolution: 2.3  Å
Similar Structures:
Biological Unit for 3WCK: monomeric; determined by author and by software (PISA)
Molecular Components in 3WCK
Label Count Molecule
Protein (1 molecule)
1
Monomeric Photosenitizing Fluorescent Protein Supernova
Molecule annotation
* Click molecule labels to explore molecular sequence information.

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