6H01
Crystal structure of a domain-swapped dark-state sfGFP containing the unnatural amino acid ortho-nitrobenzyl-tyrosine (ONBY) at residue 66
Summary for 6H01
| Entry DOI | 10.2210/pdb6h01/pdb |
| Descriptor | Green fluorescent protein (2 entities in total) |
| Functional Keywords | gfp, unnatural amino acids, domain-swapped, fluorescent protein |
| Biological source | Aequorea victoria (Jellyfish) |
| Total number of polymer chains | 4 |
| Total formula weight | 119633.65 |
| Authors | Kesgin-Schaefer, S.,Tidow, H. (deposition date: 2018-07-06, release date: 2019-04-24, Last modification date: 2024-11-20) |
| Primary citation | Kesgin-Schaefer, S.,Heidemann, J.,Puchert, A.,Koelbel, K.,Yorke, B.A.,Huse, N.,Pearson, A.R.,Uetrecht, C.,Tidow, H. Crystal structure of a domain-swapped photoactivatable sfGFP variant provides evidence for GFP folding pathway. Febs J., 286:2329-2340, 2019 Cited by PubMed Abstract: Photoactivatable fluorescent proteins (PA-FPs) are a powerful non-invasive tool in high-resolution live-cell imaging. They can be converted from an inactive to an active form by light, enabling the spatial and temporal trafficking of proteins and cell dynamics. PA-FPs have been previously generated by mutating selected residues in the chromophore or in its close proximity. A new strategy to generate PA-FPs is the genetic incorporation of unnatural amino acids (UAAs) containing photocaged groups using unique suppressor tRNA/aminoacyl-tRNA synthetase pairs. We set out to develop a photoactivatable GFP variant suitable for time-resolved structural studies. Here, we report the crystal structure of superfolder GFP (sfGFP) containing the UAA ortho-nitrobenzyl-tyrosine (ONBY) at position 66 and its spectroscopic characterization. Surprisingly, the crystal structure (to 2.7 Å resolution) reveals a dimeric domain-swapped arrangement of sfGFP66ONBY with residues 1-142 of one molecule associating with residues 148-234 from another molecule. This unusual domain-swapped structure supports a previously postulated GFP folding pathway that proceeds via an equilibrium intermediate. PubMed: 30817081DOI: 10.1111/febs.14797 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.776 Å) |
Structure validation
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