The cytidine at the first anticodon position of archaeal tRNA Ile2, which decodes the isoleucine AUA codon, is modified to 2-agmatinylcytidine (agm 2C) to guarantee the fidelity of protein biosynthesis. This post-transcriptional modification is catalyzed by tRNA Ile-agm 2C synthetase (TiaS) using ATP and agmatine as substrates. Archaeoglobus fulgidus TiaS was overexpressed in Escherichia coli cells and purified. tRNA Ile2 was prepared by in vitro transcription with T7 RNA polymerase. TiaS was cocrystallized with both tRNA Ile2 and ATP by the vapour-diffusion method. The crystals of the TiaS-tRNA Ile2-ATP complex diffracted to 2.9 Å resolution using synchrotron radiation at the Photon Factory. The crystals belonged to the primitive hexagonal space group P3221, with unit-cell parameters a = b = 131.1, c = 86.6 Å. The asymmetric unit is expected to contain one TiaS-tRNA Ile2-ATP complex, with a Matthews coefficient of 2.8 Å 3 Da -1 and a solvent content of 61%.
|Number of pages||3|
|Journal||Acta Crystallographica Section F: Structural Biology and Crystallization Communications|
|Publication status||Published - Nov 2011|
All Science Journal Classification (ASJC) codes
- Structural Biology
- Condensed Matter Physics