Crystallization and preliminary X-ray analysis of the mRNA-binding domain of elongation factor SelB from Escherichia coli in complex with RNA.
In bacteria, selenocysteine (the 21st amino acid) is incorporated into proteins via machinery that includes SelB, a specific translational elongation factor. SelB binds to an mRNA hairpin called the selenocysteine-insertion sequence (SECIS) and delivers selenocysteyl-tRNA(Sec) to the ribosomal A site. The minimum C-terminal fragment (residues 478-614) of Escherichia coli SelB (SelB-WH3/4) required for SECIS binding has been overexpressed and purified. This protein was crystallized in complex with 23 nucleotides of the SECIS hairpin at 294 K using the hanging-drop vapour-diffusion method. A data set was collected to 2.3 A resolution from a single crystal at 100 K using ESRF beamline BM-30. The crystal belongs to space group C2, with unit-cell parameters a = 103.50, b = 56.51, c = 48.41 A. The asymmetric unit contains one WH3/4-domain-RNA complex. The Matthews coefficient was calculated to be 3.37 A3 Da(-1) and the solvent content was estimated to be 67.4%.
Soler
Nicolas
N
Laboratoire de Chimie et Biologie Structurales, ICSN-CNRS, 1 Avenue de la Terrasse, 91190 Gif-sur-Yvette, France.
Fourmy
Dominique
D
Yoshizawa
Satoko
S
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D017931:DNA Primers; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D012333:RNA, Messenger / Q000378:metabolism*
D001426:Bacterial Proteins; D017931:DNA Primers; D029968:Escherichia coli Proteins; D012333:RNA, Messenger; C061878:SelB protein, Bacteria