Crystallisation of the glycyl-tRNA synthetase from Thermus thermophilus and initial crystallographic data.
The glycyl-tRNA synthetase from Thermus thermophilus is a dimer of molecular mass 115 kDa, which has been crystallised using the vapour diffusion method from 5 to 7% polyethylene glycol 6000, 0.8 to 1.4 M NaCl at protein concentrations of 2 to 8 mg/ml. Nucleation is carried out at 4 degrees C and crystals are subsequently transferred to 15 degrees C to maximise growth. Crystals are truncated rhombohedra measuring on average 0.4 mm x 0.4 mm x 0.2 mm, which appear within a few days and reach full size in one to two months. GlyRS crystallises in two closely related space groups, P2(1)2(1)2(1) and C2,2,2(1), both with the same cell a = 125 A, b = 254 A, c = 104 A. Crystal packing in P2(1)2(1)2(1) is strongly C-centred. The crystals have VM = 3.6 A3/Da and a solvent content of 61%, with one dimer in the asymmetric unit in C2,2,2(1) and two dimers in P2(1)2(1)2(1). The best native data extend to 2.9 A in C2,2,2(1) and are 90.6% complete with an R-factor between symmetry-related reflections of 10.0%. The structure has been solved by multiple isomorphous replacement and model building is in progress.
Logan
D T
DT
UPR de Biologie Structurale, Strasbourg, France.
Cura
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Touzel
J P
JP
Kern
D
D
Moras
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D
D003460:Crystallization; D018360:Crystallography, X-Ray; D006032:Glycine-tRNA Ligase / Q000737:chemistry*; D015394:Molecular Structure; D016963:Thermus thermophilus / Q000201:enzymology*
D006032:Glycine-tRNA Ligase