1ioz

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spinqualitylabelsall models 1ioz, resolution 2.0Å Show:Asymmetric Unit Biological Assembly Drag the structure with the mouse to rotate   Export Animated Image

CRYSTAL STRUCTURE OF THE C-HA-RAS PROTEIN PREPARED BY THE CELL-FREE SYNTHESIS

Contents 1 Overview 2 Disease 3 About this Structure 4 Reference

Overview

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.

Disease

Known diseases associated with this structure: Bladder cancer, somatic OMIM:[190020], Costello syndrome OMIM:[190020], Thyroid carcinoma, follicular, somatic OMIM:[190020]

About this Structure

1IOZ is a Single protein structure of sequence from Homo sapiens with GDP as ligand. Full crystallographic information is available from OCA.

Reference

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

Page seeded by OCA on Mon Nov 12 17:32:44 2007