Open AccessPurification, crystallization and preliminary X‐ray analysis of the catalytic domain of the Escherichia coli tRNase colicin D
Author(s) -
Takahashi Kazutoshi,
Ogawa Tetsuhiro,
Hidaka Makoto,
Ohsawa Kanju,
Masaki Haruhiko,
Yajima Shunsuke
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105039679
Subject(s) - escherichia coli , crystallization , colicin , crystallography , tris , chemistry , resolution (logic) , transfer rna , stereochemistry , rna , biochemistry , organic chemistry , artificial intelligence , computer science , gene
The tRNase domain of colicin D, which cleaves only tRNA Arg s at the 3′ side of their anticodon loops, has been expressed in Escherichia coli with its inhibitor protein and purified to a form free from the inhibitor using a low‐pH buffer. Crystals were obtained by the hanging‐drop vapour‐diffusion method at 278 K from a buffer containing 100 m M Tris–HCl pH 8.5, 22% PEG MME 2000 and 1 m M nickel(II) chloride. Diffraction data to 1.05 Å resolution were collected at BL41XU, SPring‐8. The crystals belong to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 34.7, b = 65.5, c = 96.5 Å.