title: Expression, crystallization and preliminary crystallographic studies of a novel bifunctional N-acetylglutamate synthase/kinase from Xanthomonas campestris homologous to vertebrate N-acetylglutamate synthase.
abstract: 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.
authors.affiliation: Children's National Medical Center, The George Washington University, 111 Michigan Avenue, Washington, DC 20010, USA. dshi@cnmcresearch.org
authors.forename: Dashuang
authors.initials: D
authors.lastname: Shi
authors.forename: Ljubica
authors.initials: L
authors.lastname: Caldovic
authors.forename: Zhongmin
authors.initials: Z
authors.lastname: Jin
authors.forename: Xiaolin
authors.initials: X
authors.lastname: Yu
authors.forename: Qiuhao
authors.initials: Q
authors.lastname: Qu
authors.forename: Lauren
authors.initials: L
authors.lastname: Roth
authors.forename: Hiroki
authors.initials: H
authors.lastname: Morizono
authors.forename: Yetrib
authors.initials: Y
authors.lastname: Hathout
authors.forename: Norma M
authors.initials: NM
authors.lastname: Allewell
authors.forename: Mendel
authors.initials: M
authors.lastname: Tuchman
mesh_terms: 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*
chemical_list: D009097:Multienzyme Complexes; D011994:Recombinant Proteins; D012645:Selenomethionine; D051046:Amino-Acid N-Acetyltransferase; D017851:Phosphotransferases (Carboxyl Group Acceptor); C018721:acetylglutamate kinase