Vandavasi Venugopal, Taylor-Creel Kasey, McFeeters Robert L, Coates Leighton, McFeeters Hana
Biology and Soft Matter Division, Oak Ridge National Laboratory, PO Box 2008, Oak Ridge, TN 37831, USA.
Department of Chemistry, University of Alabama in Huntsville, 301 Sparkman Drive, Huntsville, AL 35899, USA.
Acta Crystallogr F Struct Biol Commun. 2014 Jul;70(Pt 7):872-7. doi: 10.1107/S2053230X14009893. Epub 2014 Jun 18.
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.
来自致病性细菌鼠伤寒沙门氏菌的肽基-tRNA水解酶（Pth；EC 3.1.1.29）已被克隆，在大肠杆菌中表达并结晶以进行X射线分析。晶体通过悬滴气相扩散法，以由0.03 M柠檬酸、0.05 M双三羟甲基丙烷、1%甘油、3%蔗糖、25%聚乙二醇6000（pH 7.6）组成的储液生长。晶体用于以1.6 Å分辨率获得天然蛋白质的三维结构。该结构通过分子置换法确定，置换的晶体学数据来自空间群为P2₁2₁2₁的晶胞参数a = 62.1、b = 64.9、c = 110.5 Å，α = β = γ = 90°。晶体学晶格的不对称单元由酶分子的两个拷贝组成，溶剂含量为51%，对应于马修斯系数为2.02 ų Da⁻¹。所报道的结构坐标为针对新型小分子Pth1抑制剂和潜在抗菌药物开发的计算和结构导向研究奠定了基础。
