Open AccessCrystallization and preliminary X‐ray crystallographic analysis of peptide deformylase (PDF) from Bacillus cereus in ligand‐free and actinonin‐bound forms
Author(s) -
Park Joon Kyu,
Moon Jin Ho,
Kim JaeHong,
Kim Eunice EunKyeong
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309104032440
Subject(s) - bacillus cereus , escherichia coli , crystallization , ligand (biochemistry) , cereus , crystallography , resolution (logic) , bacteria , chemistry , stereochemistry , peptide , biochemistry , biology , organic chemistry , gene , receptor , genetics , artificial intelligence , computer science
In bacteria, protein expression initiates with an N ‐formyl group and this needs to be removed in order to ensure proper bacterial growth. These formylation and deformylation processes are unique to eubacteria; therefore, inhibition of these would provide a novel antibacterial therapy. Deformylation is carried out by peptide deformylase (PDF). PDF from Bacillus cereus , one of the major pathogenic bacteria, was cloned into expression plasmid pET‐28a (Novagen), overexpressed in Escherichia coli BL21 (DE3) and purified to high quality. Crystals have been obtained of both ligand‐free PDF and PDF to which actinonin, a highly potent naturally occurring inhibitor, is bound. Both crystals belong to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 42.72, b = 44.04, c = 85.19 Å and a = 41.31, b = 44.56, c = 84.47 Å, respectively. Diffraction data were collected to 1.7 Å resolution for the inhibitor‐free crystals and to 2.0 Å resolution for the actinonin‐bound crystals.