Open AccessCrystallization and preliminary X‐ray analysis of the mRNA‐binding domain of elongation factor SelB in complex with RNA
Author(s) -
Rasubala Linda,
Fourmy Dominique,
Ose Toyoyuki,
Kohda Daisuke,
Maenaka Katsumi,
Yoshizawa Satoko
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105003611
Subject(s) - elongation , crystallization , rna , messenger rna , elongation factor , rna binding protein , crystallography , biophysics , chemistry , biology , biochemistry , materials science , gene , ribosome , metallurgy , organic chemistry , ultimate tensile strength
In bacteria, the selenocysteine‐specific elongation factor SelB is necessary for incorporation of selenocysteine, the 21st amino acid, into proteins by the ribosome. SelB binds to an mRNA hairpin formed by the selenocysteine‐insertion sequence (SECIS) and delivers selenocysteyl‐tRNA (Sec‐tRNA Sec ) at the ribosomal A site. The minimum fragment (residues 512–634) of Moorella thermoacetica SelB (SelB‐M) required for mRNA binding has been overexpressed and purified. The complex of SelB‐M with 23 nucleotides of the SECIS mRNA hairpin was crystallized at 293 K using the hanging‐drop vapour‐diffusion or oil‐batch methods. The crystals diffract to 2.3 Å resolution using SPring‐8 BL41XU and belong to the space group P 2 1 2 1 2, with unit‐cell parameters a = 81.69, b = 169.58, c = 71.69 Å.