PMID:10393305
| Citation |
Gil, F, Ramón-Maiques, S, Marina, A, Fita, I and Rubio, V (1999) N-Acetyl-L-glutamate kinase from Escherichia coli: cloning of the gene, purification and crystallization of the recombinant enzyme and preliminary X-ray analysis of the free and ligand-bound forms. Acta Crystallogr. D Biol. Crystallogr. 55:1350-2 |
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| Abstract |
The gene for Escherichia coli N-acetyl-L-glutamate kinase (NAGK) was cloned in a plasmid and expressed in E. coli, allowing enzyme purification in three steps. NAGK exhibits high specific activity (1.1 micromol s-1 mg-1), lacks Met1 and forms dimers (shown by cross-linking). Crystals of unliganded NAGK diffract to 2 A and belong to space group P6122 or its enantiomorph P6522 (unit-cell parameters a = b = 78.6, c = 278.0 A) with two monomers in the asymmetric unit. Crystals of NAGK with acetylglutamate and the ATP analogue AMPPNP diffract to 1.8 A and belong to space group C2221 (unit-cell parameters a = 60.0, b = 71.9, c = 107.4 A), with one monomer in the asymmetric unit. NAGK crystallization will allow the determination of proposed structural similarities to carbamate kinase. |
| Links | |
| Keywords |
Base Sequence; Cloning, Molecular; Crystallization; Crystallography, X-Ray; DNA Primers; Escherichia coli/enzymology; Phosphotransferases (Carboxyl Group Acceptor)/chemistry; Phosphotransferases (Carboxyl Group Acceptor)/genetics; Phosphotransferases (Carboxyl Group Acceptor)/isolation & purification; Protein Conformation; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification |
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