5OXC
Structure of Cerulean Fluorescent Protein at 1.02 Angstrom resolution
Summary for 5OXC
| Entry DOI | 10.2210/pdb5oxc/pdb |
| Descriptor | Green fluorescent protein (2 entities in total) |
| Functional Keywords | fluorescent protein, tryptophan-based chromophore, cerulean, hydrogen atoms |
| Biological source | Aequorea victoria (Jellyfish) |
| Total number of polymer chains | 1 |
| Total formula weight | 26791.25 |
| Authors | Gotthard, G.,von Stetten, D.,Clavel, D.,Noirclerc-Savoye, M.,Royant, A. (deposition date: 2017-09-06, release date: 2017-11-29, Last modification date: 2024-10-16) |
| Primary citation | Gotthard, G.,von Stetten, D.,Clavel, D.,Noirclerc-Savoye, M.,Royant, A. Chromophore Isomer Stabilization Is Critical to the Efficient Fluorescence of Cyan Fluorescent Proteins. Biochemistry, 56:6418-6422, 2017 Cited by PubMed Abstract: ECFP, the first usable cyan fluorescent protein (CFP), was obtained by adapting the tyrosine-based chromophore environment in green fluorescent protein to that of a tryptophan-based one. This first-generation CFP was superseded by the popular Cerulean, CyPet, and SCFP3A that were engineered by rational and random mutagenesis, yet the latter CFPs still exhibit suboptimal properties of pH sensitivity and reversible photobleaching behavior. These flaws were serendipitously corrected in the third-generation CFP mTurquoise and its successors without an obvious rationale. We show here that the evolution process had unexpectedly remodeled the chromophore environment in second-generation CFPs so they would accommodate a different isomer, whose formation is favored by acidic pH or light irradiation and which emits fluorescence much less efficiently. Our results illustrate how fluorescent protein engineering based solely on fluorescence efficiency optimization may affect other photophysical or physicochemical parameters and provide novel insights into the rational evolution of fluorescent proteins with a tryptophan-based chromophore. PubMed: 29148725DOI: 10.1021/acs.biochem.7b01088 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.02 Å) |
Structure validation
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