title: Recombinant production, crystallization and X-ray crystallographic structure determination of peptidyl-tRNA hydrolase from Salmonella typhimurium.
abstract: Peptidyl-tRNA hydrolase (Pth; EC 3.1.1.29) from the pathogenic bacterium Salmonella typhimurium has been cloned, expressed in Escherichia coli and crystallized for X-ray analysis. Crystals were grown using hanging-drop vapor diffusion against a reservoir solution consisting of 0.03 M citric acid, 0.05 M bis-tris propane, 1% glycerol, 3% sucrose, 25% PEG 6000 pH 7.6. Crystals were used to obtain the three-dimensional structure of the native protein at 1.6 Å resolution. The structure was determined by molecular replacement of the crystallographic data processed in space group P2₁2₁2₁ with unit-cell parameters a=62.1, b=64.9, c=110.5 Å, α=β=γ=90°. The asymmetric unit of the crystallographic lattice was composed of two copies of the enzyme molecule with a 51% solvent fraction, corresponding to a Matthews coefficient of 2.02 Å3 Da(-1). The structural coordinates reported serve as a foundation for computational and structure-guided efforts towards novel small-molecule Pth1 inhibitors and potential antibacterial development.
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, PO Box 2008, Oak Ridge, TN 37831, USA.
authors.forename: Venugopal
authors.initials: V
authors.lastname: Vandavasi
authors.affiliation: Department of Chemistry, University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA.
authors.forename: Kasey
authors.initials: K
authors.lastname: Taylor-Creel
authors.affiliation: Department of Chemistry, University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA.
authors.forename: Robert L
authors.initials: RL
authors.lastname: McFeeters
authors.affiliation: Biology and Soft Matter Division, Oak Ridge National Laboratory, PO Box 2008, Oak Ridge, TN 37831, USA.
authors.forename: Leighton
authors.initials: L
authors.lastname: Coates
authors.affiliation: Department of Chemistry, University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA.
authors.forename: Hana
authors.initials: H
authors.lastname: McFeeters
mesh_terms: D002265:Carboxylic Ester Hydrolases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D005990:Glycerol / Q000737:chemistry; D006863:Hydrogen-Ion Concentration; D008958:Models, Molecular; D011092:Polyethylene Glycols / Q000737:chemistry; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012486:Salmonella typhimurium / Q000737:chemistry* / Q000201:enzymology; D040681:Structural Homology, Protein; D013395:Sucrose / Q000737:chemistry
chemical_list: D011994:Recombinant Proteins; D011092:Polyethylene Glycols; D013395:Sucrose; D002265:Carboxylic Ester Hydrolases; C019674:aminoacyl-tRNA hydrolase; D005990:Glycerol
keywords: Salmonella typhimurium; peptidyl-tRNA; peptidyl-tRNA hydrolase 1