3U8C
Crystal structure of monomeric reversibly photoswitchable red fluorescent protein rsTagRFP in the ON state
Summary for 3U8C
| Entry DOI | 10.2210/pdb3u8c/pdb |
| Related | 3M22 3U8A |
| Descriptor | Fluorescent protein rsTagRFP, GLYCEROL (3 entities in total) |
| Functional Keywords | beta barrel, fluorescent protein, light induced reversible photoswitching, reversibly photoswitchable fluorescent protein, cis-trans isomerization |
| Biological source | synthetic construct (artificial gene) |
| Total number of polymer chains | 4 |
| Total formula weight | 110649.91 |
| Authors | Pletnev, S. (deposition date: 2011-10-16, release date: 2012-02-22, Last modification date: 2025-03-26) |
| Primary citation | Pletnev, S.,Subach, F.V.,Dauter, Z.,Wlodawer, A.,Verkhusha, V.V. A structural basis for reversible photoswitching of absorbance spectra in red fluorescent protein rsTagRFP. J.Mol.Biol., 417:144-151, 2012 Cited by PubMed Abstract: rsTagRFP is the first monomeric red fluorescent protein (FP) with reversibly photoswitchable absorbance spectra. The switching is realized by irradiation of rsTagRFP with blue (440 nm) and yellow (567 nm) light, turning the protein fluorescence ON and OFF, respectively. It is perhaps the most useful probe in this color class that has yet been reported. Because of the photoswitchable absorbance, rsTagRFP can be used as an acceptor in photochromic Förster resonance energy transfer. Yellow FPs, YPet and mVenus, are demonstrated to be excellent photochromic Förster resonance energy transfer donors for the rsTagRFP acceptor in its fusion constructs. Analysis of X-ray structures has shown that photoswitching of rsTagRFP is accompanied by cis-trans isomerization and protonation/deprotonation of the chromophore, with the deprotonated cis- and protonated trans-isomers corresponding to its ON and OFF states, respectively. Unlike in other photoswitchable FPs, both conformers of rsTagRFP chromophore are essentially coplanar. Two other peculiarities of the rsTagRFP chromophore are an essentially hydrophobic environment of its p-hydroxyphenyl site and the absence of direct hydrogen bonding between this moiety and the protein scaffold. The influence of the immediate environment on rsTagRFP chromophore was probed by site-directed mutagenesis. Residues Glu145 and His197 were found to participate in protonation/deprotonation of the chromophore accompanying the photoswitching of rsTagRFP fluorescence, whereas residues Met160 and Leu174 were shown to spatially restrict chromophore isomerization, favoring its radiative decay. PubMed: 22310052DOI: 10.1016/j.jmb.2012.01.044 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.188 Å) |
Structure validation
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