Expression, crystallization and preliminary crystallographic studies of a novel bifunctional N-acetylglutamate synthase/kinase from Xanthomonas campestris homologous to vertebrate N-acetylglutamate synthase.
A novel N-acetylglutamate synthase/kinase bifunctional enzyme of arginine biosynthesis that was homologous to vertebrate N-acetylglutamate synthases was identified in Xanthomonas campestris. The protein was overexpressed, purified and crystallized. The crystals belong to the hexagonal space group P6(2)22, with unit-cell parameters a = b = 134.60, c = 192.11 A, and diffract to about 3.0 A resolution. Selenomethionine-substituted recombinant protein was produced and selenomethionine substitution was verified by mass spectroscopy. Multiple anomalous dispersion (MAD) data were collected at three wavelengths at SER-CAT, Advanced Photon Source, Argonne National Laboratory. Structure determination is under way using the MAD phasing method.
Shi
Dashuang
D
Children's National Medical Center, The George Washington University, 111 Michigan Avenue, Washington, DC 20010, USA. dshi@cnmcresearch.org
Caldovic
Ljubica
L
Jin
Zhongmin
Z
Yu
Xiaolin
X
Qu
Qiuhao
Q
Roth
Lauren
L
Morizono
Hiroki
H
Hathout
Yetrib
Y
Allewell
Norma M
NM
Tuchman
Mendel
M
D051046:Amino-Acid N-Acetyltransferase / Q000096:biosynthesis / Q000737:chemistry*; D003460:Crystallization; D018360:Crystallography, X-Ray; D009097:Multienzyme Complexes / Q000096:biosynthesis / Q000737:chemistry*; D017851:Phosphotransferases (Carboxyl Group Acceptor) / Q000096:biosynthesis / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis; D012645:Selenomethionine / Q000378:metabolism; D016959:Xanthomonas campestris / Q000201:enzymology*
D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D012645:Selenomethionine; D051046:Amino-Acid N-Acetyltransferase; D017851:Phosphotransferases (Carboxyl Group Acceptor); C018721:acetylglutamate kinase