Crystallization and preliminary X-ray diffraction analysis of an archaeal tRNA-modification enzyme, TiaS, complexed with tRNA Ile2 and ATP

Takuo Osawa, Hideko Inanaga, Satoshi Kimura, Naohiro Terasaka, Tsutomu Suzuki, Tomoyuki Numata

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

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%.

Original languageEnglish
Pages (from-to)1414-1416
Number of pages3
JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
Volume67
Issue number11
DOIs
Publication statusPublished - Nov 2011
Externally publishedYes

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Genetics
  • Condensed Matter Physics

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