Expression, purification and preliminary X-ray characterization of N-acetyl-gamma-glutamyl-phosphate reductase from Thermus thermophilus HB8.
N-Acetyl-gamma-glutamyl-phosphate reductase (AGPR) catalyses the NADPH-dependent reduction of N-acetyl-gamma-glutamyl phosphate to give the N-acetylglutamic semialdehyde. A recombinant form of AGPR from Thermus thermophilus HB8 has been crystallized by the hanging-drop vapour-diffusion technique using PEG 4000 as a precipitating agent. The crystals grew as colourless prisms, with unit-cell parameters a = b = 90.9, c = 139.5 A, alpha = beta = 90, gamma = 120 degrees. The crystals belong to the hexagonal space group P6(2)22 or P6(4)22 and are most likely to contain a monomer in the asymmetric unit, with a V(M) value of 2.19 A(3) Da(-1). The crystals diffract to a resolution of 2.2 A at beamline BL44B2 of SPring-8.
Goto
Masaru
M
Department of Chemistry, Graduate School of Science, Osaka City University, Sumiyoshi-ku, Osaka 558-8585, Japan.
Agari
Yoshihiro
Y
Omi
Rie
R
Miyahara
Ikuko
I
Hirotsu
Ken
K
D000445:Aldehyde Oxidoreductases / Q000096:biosynthesis / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D003460:Crystallization / Q000379:methods; D018360:Crystallography, X-Ray; D017931:DNA Primers / Q000235:genetics; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D016963:Thermus thermophilus / Q000201:enzymology* / Q000235:genetics
D017931:DNA Primers; D011994:Recombinant Proteins; D000445:Aldehyde Oxidoreductases; C018722:N-acetyl-gamma-glutamyl-phosphate reductase