Expression, purification and preliminary X-ray characterization of CTP synthetase from Thermus thermophilus HB8.
A recombinant form of the CTP synthetase from Thermus thermophilus HB8 (tCTPs) was grown as colourless crystals by the hanging-drop vapour-diffusion technique using ammonium sulfate or sodium citrate as a precipitating agent. The crystals belong to space group I222, with unit-cell parameters a = 88.2, b = 118.9, c = 142.7 A, alpha = beta = gamma = 90 degrees, and are most likely to contain a monomer in the asymmetric unit with a V(M) value of 3.07 A(3) Da(-1). The crystals obtained from ammonium sulfate and sodium citrate solutions diffract X-rays to a resolution of 2.25 A using synchrotron X-ray sources and to a resolution of 2.35 A using Cu Kalpha X-rays from a rotating-anode generator.
Goto
Masaru
M
Department of Chemistry, Graduate School of Science, Osaka City University, Japan.
Omi
Rie
R
Hoseki
Jun
J
Nakagawa
Noriko
N
Miyahara
Ikuko
I
Hirotsu
Ken
K
D019731:Carbon-Nitrogen Ligases / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000378:metabolism; D011994:Recombinant Proteins; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016963:Thermus thermophilus / Q000201:enzymology*
D011994:Recombinant Proteins; D019731:Carbon-Nitrogen Ligases; C019951:CTP synthetase