TY - JOUR
T1 - Crystallization and preliminary X-ray crystallographic analysis of a bacterial Asn-transamidosome
AU - Suzuki, Tateki
AU - Yamashita, Keitaro
AU - Tanaka, Yoshikazu
AU - Tanaka, Isao
AU - Yao, Min
PY - 2014/6
Y1 - 2014/6
N2 - Most canonical aminoacyl-tRNAs are synthesized directly by their cognate aminoacyl-tRNA synthetases (aaRSs), but glutaminyl-tRNAGln and asparaginyl-tRNAAsn 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 tRNAAsn. The crystal of the Asn-transamidosome belonged to space group P21, 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 tRNAAsns, indicating that the construction of the current Asn-transamidosome differs from that of Thermus thermophilus.
AB - Most canonical aminoacyl-tRNAs are synthesized directly by their cognate aminoacyl-tRNA synthetases (aaRSs), but glutaminyl-tRNAGln and asparaginyl-tRNAAsn 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 tRNAAsn. The crystal of the Asn-transamidosome belonged to space group P21, 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 tRNAAsns, indicating that the construction of the current Asn-transamidosome differs from that of Thermus thermophilus.
KW - AspRS
KW - GatCAB
KW - Pseudomonas aeruginosa
KW - aminoacyl-tRNA
KW - transamidosome
UR - http://www.scopus.com/inward/record.url?scp=84905487458&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84905487458&partnerID=8YFLogxK
U2 - 10.1107/S2053230X14007274
DO - 10.1107/S2053230X14007274
M3 - Article
C2 - 24915095
AN - SCOPUS:84905487458
SN - 1744-3091
VL - 70
SP - 790
EP - 793
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
IS - 6
ER -