Open AccessImproved expression, purification and crystallization of a putative N ‐acetyl‐γ‐glutamyl‐phosphate reductase from rice ( Oryza sativa )
Author(s) -
MiuraOhnuma Jun,
aka Tsuyoshi,
Katoh Shizue,
Murata Katsuyoshi,
Kita Akiko,
Miki Kunio,
Katoh Etsuko
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309105035384
Subject(s) - oryza sativa , crystallization , phosphate , chemistry , biochemistry , rice plant , reductase , enzyme , biology , agronomy , gene , organic chemistry
N ‐Acetyl‐γ‐glutamyl‐phosphate reductase (AGPR) catalyzes the third step in an eight‐step arginine‐biosynthetic pathway that starts with glutamate. This enzyme converts N ‐acetyl‐γ‐glutamyl phosphate to N ‐acetylglutamate‐γ‐semialdehyde by an NADPH‐dependent reductive dephosphorylation. AGPR from Oryza sativa (OsAGPR) was expressed in Escherichia coli at 291 K as a soluble fusion protein with an upstream thioredoxin‐hexahistidine [Trx‐(His) 6 ] extension. OsAGPR(Ala50–Pro366) was purified and crystals were obtained using the sitting‐drop vapour‐diffusion method at 293 K and diffract X‐rays to at least 1.8 Å resolution. They belong to the hexagonal space group P 6 1 , with unit‐cell parameters a = 86.11, c = 316.3 Å.