7XHS
Crystal structure of CipA crystal produced by cell-free protein synthesis
Summary for 7XHS
| Entry DOI | 10.2210/pdb7xhs/pdb |
| Descriptor | Cro/Cl family transcriptional regulator (2 entities in total) |
| Functional Keywords | in cell crystal, crystalline inclusion protein |
| Biological source | Photorhabdus |
| Total number of polymer chains | 1 |
| Total formula weight | 11734.30 |
| Authors | Abe, S.,Tanaka, J.,Kojima, M.,Kanamaru, S.,Yamashita, K.,Hirata, K.,Ueno, T. (deposition date: 2022-04-10, release date: 2023-02-01, Last modification date: 2024-05-29) |
| Primary citation | Abe, S.,Tanaka, J.,Kojima, M.,Kanamaru, S.,Hirata, K.,Yamashita, K.,Kobayashi, A.,Ueno, T. Cell-free protein crystallization for nanocrystal structure determination. Sci Rep, 12:16031-16031, 2022 Cited by PubMed Abstract: In-cell protein crystallization (ICPC) has been investigated as a technique to support the advancement of structural biology because it does not require protein purification and a complicated crystallization process. However, only a few protein structures have been reported because these crystals formed incidentally in living cells and are insufficient in size and quality for structure analysis. Here, we have developed a cell-free protein crystallization (CFPC) method, which involves direct protein crystallization using cell-free protein synthesis. We have succeeded in crystallization and structure determination of nano-sized polyhedra crystal (PhC) at a high resolution of 1.80 Å. Furthermore, nanocrystals were synthesized at a reaction scale of only 20 μL using the dialysis method, enabling structural analysis at a resolution of 1.95 Å. To further demonstrate the potential of CFPC, we attempted to determine the structure of crystalline inclusion protein A (CipA), whose structure had not yet been determined. We added chemical reagents as a twinning inhibitor to the CFPC solution, which enabled us to determine the structure of CipA at 2.11 Å resolution. This technology greatly expands the high-throughput structure determination method of unstable, low-yield, fusion, and substrate-biding proteins that have been difficult to analyze with conventional methods. PubMed: 36192567DOI: 10.1038/s41598-022-19681-9 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.11 Å) |
Structure validation
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