6XY1
Crystal structure of a de novo designed parallel four-helix coiled coil, 4-KE-4.
Summary for 6XY1
| Entry DOI | 10.2210/pdb6xy1/pdb |
| Descriptor | 4-KE-4 (2 entities in total) |
| Functional Keywords | coiled coil, tetramer, parallel, de novo protein |
| Biological source | synthetic construct |
| Total number of polymer chains | 4 |
| Total formula weight | 14773.52 |
| Authors | Edgell, C.L.,Savery, N.J.,Woolfson, D.N. (deposition date: 2020-01-29, release date: 2020-03-18, Last modification date: 2024-11-06) |
| Primary citation | Edgell, C.L.,Savery, N.J.,Woolfson, D.N. RobustDe Novo-Designed Homotetrameric Coiled Coils. Biochemistry, 59:1087-1092, 2020 Cited by PubMed Abstract: -designed protein domains are increasingly being applied in biotechnology, cell biology, and synthetic biology. Therefore, it is imperative that these proteins be robust to superficial changes; i.e., small changes to their amino acid sequences should not cause gross structural changes. In turn, this allows properties such as stability and solubility to be tuned without affecting structural attributes like tertiary fold and quaternary interactions. Reliably designed proteins with predictable behaviors may then be used as scaffolds to incorporate function, e.g., through the introduction of features for small-molecule, metal, or macromolecular binding, and enzyme-like active sites. Generally, achieving this requires the starting protein fold to be well understood. Herein, we focus on designing α-helical coiled coils, which are well studied, widespread, and often direct protein-protein interactions in natural systems. Our initial investigations reveal that a previously designed parallel, homotetrameric coiled coil, CC-Tet, is not robust to sequence changes that were anticipated to maintain its structure. Instead, the alterations switch the oligomeric state from tetramer to trimer. To improve the robustness of designed homotetramers, additional sequences based on CC-Tet were produced and characterized in solution and by X-ray crystallography. Of these updated sequences, one is robust to truncation and to changes in surface electrostatics; we call this CC-Tet*. Variants of the general CC-Tet* design provide a set of homotetrameric coiled coils with unfolding temperatures in the range from 40 to >95 °C. We anticipate that these will be of use in applications requiring robust and well-defined tetramerization domains. PubMed: 32133841DOI: 10.1021/acs.biochem.0c00082 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.5 Å) |
Structure validation
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