Crystallization and preliminary X-ray crystallographic analysis of a bacterial Asn-transamidosome.

ABSTRACT: 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 P2?, with unit-cell parameters a=93.3, b=186.0, c=287.8?Å, ?=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 tRNAAsns, indicating that the construction of the current Asn-transamidosome differs from that of Thermus thermophilus.

SUBMITTER: Suzuki T 

PROVIDER: S-EPMC4051539 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature