Premium
Crystallization and preliminary X‐ray crystallographic analysis of a bacterial Asn‐transamidosome
Author(s) -
Suzuki Tateki,
Yamashita Keitaro,
Tanaka Yoshikazu,
Tanaka Isao,
Yao Min
Publication year - 2014
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 37
ISSN - 2053-230X
DOI - 10.1107/s2053230x14007274
Subject(s) - thermus thermophilus , transfer rna , dimer , crystallization , crystallography , ribonucleoprotein , chemistry , stereochemistry , biochemistry , rna , escherichia coli , organic chemistry , gene
Most canonical aminoacyl‐tRNAs are synthesized directly by their cognate aminoacyl‐tRNA synthetases (aaRSs), but glutaminyl‐tRNA Gln and asparaginyl‐tRNA Asn are synthesized indirectly by two‐step processes. These processes are catalyzed by the transamidosome, a large ribonucleoprotein particle composed of GatA, GatB, GatC, aaRS and tRNA. In this study, the Asn‐transamidosome from Pseudomonas aeruginosa was reconstructed and crystallized by mixing purified GatCAB complex, AspRS and tRNA Asn . The crystal of the Asn‐transamidosome belonged to space group P 2 1 , with unit‐cell parameters a = 93.3, b = 186.0, c = 287.8 Å, β = 93.3°, and diffracted to 3.73 Å resolution. Preliminary X‐ray crystallographic analysis showed that the asymmetric unit contained two Asn‐transamidosomes, each composed of two GatCABs, one AspRS dimer and two tRNA Asn s, indicating that the construction of the current Asn‐transamidosome differs from that of Thermus thermophilus .