2ZXV

Crystal structure of putative acetyltransferase from T. thermophilus HB8

Summary for 2ZXV

• Entry DOI: 10.2210/pdb2zxv/pdb
• Related: 2Z0Z 2Z10 2Z11
• Descriptor: Putative uncharacterized protein TTHA1799 (2 entities in total)
• Functional Keywords: alpha/beta protein, iy-substitution, structural genomics, nppsfa, national project on protein structural and functional analyses, riken structural genomics/proteomics initiative, rsgi, transferase
• Biological source: Thermus thermophilus
• Total number of polymer chains: 4
• Total formula weight: 89153.87

Authors

Murayama, K.,Kato-Murayama, M.,Terada, T.,Kuramitsu, S.,Shirouzu, M.,Yokoyama, S.,RIKEN Structural Genomics/Proteomics Initiative (RSGI) (deposition date: 2009-01-08, release date: 2009-02-17, Last modification date: 2024-11-20)

Primary citation

Sakamoto, K.,Murayama, K.,Oki, K.,Iraha, F.,Kato-Murayama, M.,Takahashi, M.,Ohtake, K.,Kobayashi, T.,Kuramitsu, S.,Shirouzu, M.,Yokoyama, S.
Genetic Encoding of 3-Iodo-l-Tyrosine in Escherichia coli for Single-Wavelength Anomalous Dispersion Phasing in Protein Crystallography
Structure, 17:335-344, 2009

PubMed Abstract: We developed an Escherichia coli cell-based system to generate proteins containing 3-iodo-L-tyrosine at desired sites, and we used this system for structure determination by single-wavelength anomalous dispersion (SAD) phasing with the strong iodine signal. Tyrosyl-tRNA synthetase from Methanocaldococcus jannaschii was engineered to specifically recognize 3-iodo-L-tyrosine. The 1.7 A crystal structure of the engineered variant, iodoTyrRS-mj, bound with 3-iodo-L-tyrosine revealed the structural basis underlying the strict specificity for this nonnatural substrate; the iodine moiety makes van der Waals contacts with 5 residues at the binding pocket. E. coli cells expressing iodoTyrRS-mj and the suppressor tRNA were used to incorporate 3-iodo-L-tyrosine site specifically into the ribosomal protein N-acetyltransferase from Thermus thermophilus. The crystal structure of this enzyme with iodotyrosine was determined at 1.8 and 2.2 Angstroms resolutions by SAD phasing at CuK alpha and CrK alpha wavelengths, respectively. The native structure, determined by molecular replacement, revealed no significant structural distortion caused by iodotyrosine incorporation.

PubMed: 19278648
DOI: 10.1016/j.str.2009.01.008

Experimental method

X-RAY DIFFRACTION (2.3 Å)