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 P2?2?2? with unit-cell parameters a=62.1, b=64.9, c=110.5?Å, ?=?=?=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.

SUBMITTER: Vandavasi V 

PROVIDER: S-EPMC4089523 | biostudies-literature | 2014 Jul

REPOSITORIES: biostudies-literature