Another application is to develop a protein–activity actuator using Dronpa mutants [43•]. With off-photoswitching, beta strand 7 near the chromophore becomes flexible. This strand forms part of the cross-dimer interface in the tetrameric parent, and so it is reasonable to expect that off-photoswitching could affect the capability of Dronpa to oligomerize. Indeed, in the dark, Dronpa Lys145Asn is tetrameric, whereas cyan illumination induced redistribution from tetrameric toward monomeric species. On the basis of this light-dependent interaction, a fluorescent light-inducible

protein (FLiPs) design was created, in which Dronpa Lys145Asn domain is fused to both termini of an enzyme of interest, where the termini straddle the enzyme active site. In the dark, the Dronpa www.selleckchem.com/HDAC.html Lys145Asn domains tetramerize and cage the protein, but light

induces Dronpa Lys145Asn dissociation and activates the protein (Figure 4b). Thus Dronpa domains can function in reversible optical control of protein activities, a type of function which had previously been assumed to exist in only other types of chromophore-containing proteins. Conveniently, the photoswitchable fluorescence of Dronpa serves as a built-in read-out of Erastin order the activity state of the target protein. It remains to be determined whether other photoswitchable FPs can also function as optical control elements. A potentially useful application of photoswitchable FPs is optical data writing and storage. Unlike photoconvertible proteins, which can create red fluorescent patterns irreversibly

created by light, photoswitchable FPs allow for multiple writing cycles [44]. from 2D data writing has been performed with Dronpa and IrisFP coated on a surface, and 3D data writing in crystals of IrisFP and other EosFP mutants [27 and 45]. Compared to other optical encoding schemes such as encoding on silver zeolite microcarriers [46], photoswitchable FPs are not as stable, and physical separation is needed to create pixels or voxels. However, they may be of utility in situations where instability or biodegradability is desirable. In the 10 years since the invention of KFP and Dronpa, photoswitchable FPs have found unique uses in the imaging of protein movements and in nanometer-scale precision localization of proteins. Just recently, a photoswitchable FP has been found to be capable of mediating control of protein activity with light, potentially expanding the uses of FPs from optical imaging to optical control. As a class of primarily artificial proteins, photoswitchable FPs continue to be the subject of protein engineering efforts as well as biophysical study to understand their unique structure and behavior.