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1ioz
From Proteopedia
| 1ioz, resolution 2.00Å () | |||||||||
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| Ligands: | |||||||||
| Domains: | H_N_K_Ras_like | ||||||||
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| Resources: | FirstGlance, OCA, PDBsum, RCSB, TOPSAN | ||||||||
| Coordinates: | save as pdb, mmCIF, xml | ||||||||
CRYSTAL STRUCTURE OF THE C-HA-RAS PROTEIN PREPARED BY THE CELL-FREE SYNTHESIS
Multi-wavelength anomalous diffraction phasing is especially useful for high-throughput structure determinations. Selenomethionine substituted proteins are commonly used for this purpose. However, the cytotoxicity of selenomethionine drastically reduces the efficiency of its incorporation in in vivo expression systems. In the present study, an improved E. coli cell-free protein synthesis system was used to incorporate selenomethionine into a protein, so that highly efficient incorporation could be achieved. A milligram quantity of selenomethionine-containing Ras was obtained using the cell-free system with dialysis. The mass spectrometry analysis showed that more than 95% of the methionine residues were substituted with selenomethionine. The crystal of this protein grew under the same conditions and had the same unit cell constants as those of the native Ras protein. The three-dimensional structure of this protein, determined by multi-wavelength anomalous diffraction phasing, was almost the same as that of the Ras protein prepared by in vivo expression. Therefore, the cell-free synthesis system could become a powerful protein expression method for high-throughput structure determinations by X-ray crystallography.
Selenomethionine incorporation into a protein by cell-free synthesis., Kigawa T, Yamaguchi-Nunokawa E, Kodama K, Matsuda T, Yabuki T, Matsuda N, Ishitani R, Nureki O, Yokoyama S, J Struct Funct Genomics. 2002;2(1):29-35. PMID:12836672
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
About this Structure
1IOZ is a 1 chain structure of sequence from Homo sapiens. Full crystallographic information is available from OCA.
Reference
- Kigawa T, Yamaguchi-Nunokawa E, Kodama K, Matsuda T, Yabuki T, Matsuda N, Ishitani R, Nureki O, Yokoyama S. Selenomethionine incorporation into a protein by cell-free synthesis. J Struct Funct Genomics. 2002;2(1):29-35. PMID:12836672
Page seeded by OCA on Tue Feb 17 22:32:37 2009
Categories: Homo sapiens | Kigawa, T. | Kodama, K. | Matsuda, T. | RSGI, RIKEN Structural Genomics/Proteomics Initiative. | Yabuki, T. | Yamaguchi-Nunokawa, E. | Cell-free protein synthesis | Gtp-binding protein | Oncogene protein | Ra | Riken structural genomics/proteomics initiative | Rsgi | Structural genomic
