TY - JOUR
T1 - Recombinant production, crystallization and X-ray crystallographic structure determination of peptidyl-tRNA hydrolase from Salmonella typhimurium
AU - Vandavasi, Venugopal
AU - Taylor-Creel, Kasey
AU - McFeeters, Robert L.
AU - Coates, Leighton
AU - McFeeters, Hana
PY - 2014/7
Y1 - 2014/7
N2 - 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 0.03M citric acid, 0.05M 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 1.6Å 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 = 110.5Å, 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 2.02Å3Da-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.
AB - 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 0.03M citric acid, 0.05M 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 1.6Å 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 = 110.5Å, 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 2.02Å3Da-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.
KW - Salmonella typhimurium
KW - peptidyl-tRNA
KW - peptidyl-tRNA hydrolase 1
UR - http://www.scopus.com/inward/record.url?scp=84905464965&partnerID=8YFLogxK
U2 - 10.1107/S2053230X14009893
DO - 10.1107/S2053230X14009893
M3 - Article
C2 - 25005080
AN - SCOPUS:84905464965
SN - 2053-230X
VL - 70
SP - 872
EP - 877
JO - Acta Crystallographica Section F:Structural Biology Communications
JF - Acta Crystallographica Section F:Structural Biology Communications
IS - 7
ER -