6SM0
Venus 66 p-Azido-L-Phenylalanin (azF) variant, dark grown
Summary for 6SM0
| Entry DOI | 10.2210/pdb6sm0/pdb |
| Descriptor | Green fluorescent protein, OXYGEN MOLECULE, ZINC ION, ... (5 entities in total) |
| Functional Keywords | venus 66 fluorescent protein, azf variant, aequoria victoria, gfp, non-canonical amino acid, 3d x-ray structure determination, fluorescent protein |
| Biological source | Aequorea victoria |
| Total number of polymer chains | 1 |
| Total formula weight | 25887.34 |
| Authors | Rizkallah, P.J.,Al Maslookhi, H.S.,Jones, D.D. (deposition date: 2019-08-21, release date: 2021-02-17, Last modification date: 2024-01-24) |
| Primary citation | Auhim, H.S.,Grigorenko, B.L.,Harris, T.K.,Aksakal, O.E.,Polyakov, I.V.,Berry, C.,Gomes, G.D.P.,Alabugin, I.V.,Rizkallah, P.J.,Nemukhin, A.V.,Jones, D.D. Stalling chromophore synthesis of the fluorescent protein Venus reveals the molecular basis of the final oxidation step. Chem Sci, 12:7735-7745, 2021 Cited by PubMed Abstract: Fluorescent proteins (FPs) have revolutionised the life sciences, but the mechanism of chromophore maturation is still not fully understood. Here we show that incorporation of a photo-responsive non-canonical amino acid within the chromophore stalls maturation of Venus, a yellow FP, at an intermediate stage; a crystal structure indicates the presence of O located above a dehydrated enolate form of the imidazolone ring, close to the strictly conserved Gly67 that occupies a twisted conformation. His148 adopts an "open" conformation so forming a channel that allows O access to the immature chromophore. Absorbance spectroscopy supported by QM/MM simulations suggests that the first oxidation step involves formation of a hydroperoxyl intermediate in conjunction with dehydrogenation of the methylene bridge. A fully conjugated mature chromophore is formed through release of HO, both and . The possibility of interrupting and photochemically restarting chromophore maturation and the mechanistic insights open up new approaches for engineering optically controlled fluorescent proteins. PubMed: 34168826DOI: 10.1039/d0sc06693a PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.91 Å) |
Structure validation
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