4KA9
Crystal structure analysis of single amino acid deletion mutations in EGFP
Summary for 4KA9
| Entry DOI | 10.2210/pdb4ka9/pdb |
| Related | 4KAG 4KEX |
| Descriptor | Green fluorescent protein, 1,2-ETHANEDIOL, 4-(2-HYDROXYETHYL)-1-PIPERAZINE ETHANESULFONIC ACID, ... (6 entities in total) |
| Functional Keywords | beta barrel, fluorescent protein, chromophore cyclisation, single amino acid deletion mutation, cyclisation |
| Biological source | Aequorea victoria (Jellyfish) |
| Total number of polymer chains | 1 |
| Total formula weight | 27873.11 |
| Authors | Arpino, J.A.J.,Rizkallah, P.J. (deposition date: 2013-04-22, release date: 2014-05-07, Last modification date: 2024-11-06) |
| Primary citation | Arpino, J.A.,Reddington, S.C.,Halliwell, L.M.,Rizkallah, P.J.,Jones, D.D. Random single amino acid deletion sampling unveils structural tolerance and the benefits of helical registry shift on GFP folding and structure. Structure, 22:889-898, 2014 Cited by PubMed Abstract: Altering a protein's backbone through amino acid deletion is a common evolutionary mutational mechanism, but is generally ignored during protein engineering primarily because its effect on the folding-structure-function relationship is difficult to predict. Using directed evolution, enhanced green fluorescent protein (EGFP) was observed to tolerate residue deletion across the breadth of the protein, particularly within short and long loops, helical elements, and at the termini of strands. A variant with G4 removed from a helix (EGFP(G4Δ)) conferred significantly higher cellular fluorescence. Folding analysis revealed that EGFP(G4Δ) retained more structure upon unfolding and refolded with almost 100% efficiency but at the expense of thermodynamic stability. The EGFP(G4Δ) structure revealed that G4 deletion caused a beneficial helical registry shift resulting in a new polar interaction network, which potentially stabilizes a cis proline peptide bond and links secondary structure elements. Thus, deletion mutations and registry shifts can enhance proteins through structural rearrangements not possible by substitution mutations alone. PubMed: 24856363DOI: 10.1016/j.str.2014.03.014 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.58 Å) |
Structure validation
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