Open AccessIsolation, crystallization and preliminary X‐ray analysis of the transamidosome, a ribonucleoprotein involved in asparagine formation
Author(s) -
Bailly Marc,
Blaise Mickael,
Lorber Bernard,
Thirup Soren,
Kern Daniel
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109015000
Subject(s) - crystallization , ribonucleoprotein , asparagine , isolation (microbiology) , crystallography , x ray , chemistry , materials science , biochemistry , biology , physics , organic chemistry , optics , bioinformatics , rna , amino acid , gene
Thermus thermophilus deprived of asparagine synthetase synthesizes Asn on tRNA Asn via a tRNA‐dependent pathway involving a nondiscriminating aspartyl‐tRNA synthetase that charges Asp onto tRNA Asn prior to conversion of the Asp to Asn by GatCAB, a tRNA‐dependent amidotransferase. This pathway also constitutes the route of Asn‐tRNA Asn formation by bacteria and archaea deprived of asparaginyl‐tRNA synthetase. The partners involved in tRNA‐dependent Asn formation in T. thermophilus assemble into a ternary complex called the transamidosome. This particule produces Asn‐tRNA Asn in the presence of free Asp, ATP and an amido‐group donor. Crystals of the transamidosome from T. thermophilus were obtained in the presence of PEG 4000 in MES–NaOH buffer pH 6.5. They belonged to the primitive monoclinic space group P 2 1 , with unit‐cell parameters a = 115.9, b = 214.0, c = 127.8 Å, β = 93.3°. A complete data set was collected to 3 Å resolution. Here, the isolation and crystallization of the transamidosome from T. thermophilus and preliminary crystallographic data are reported.