Purification, crystallization and preliminary X-ray analysis of the catalytic domain of the Escherichia coli tRNase colicin D.
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 mM Tris-HCl pH 8.5, 22% PEG MME 2000 and 1 mM nickel(II) chloride. Diffraction data to 1.05 A resolution were collected at BL41XU, SPring-8. The crystals belong to space group P2(1)2(1)2(1), with unit-cell parameters a = 34.7, b = 65.5, c = 96.5 A.
Takahashi
Kazutoshi
K
Department of Biotechnology, The University of Tokyo, Yayoi 1-1-1, Tokyo 113-8657, Japan.
Ogawa
Tetsuhiro
T
Hidaka
Makoto
M
Ohsawa
Kanju
K
Masaki
Haruhiko
H
Yajima
Shunsuke
S
D020134:Catalytic Domain; D003460:Crystallization; D018360:Crystallography, X-Ray; D004722:Endoribonucleases / Q000737:chemistry* / Q000378:metabolism; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism; D021381:Protein Transport; D012329:RNA, Bacterial / Q000737:chemistry / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000737:chemistry / Q000378:metabolism
D029968:Escherichia coli Proteins; D012329:RNA, Bacterial; D012346:RNA, Transfer, Amino Acyl; C501879:colicin D immunity protein, E coli; D004722:Endoribonucleases