PMID:9631510
| Citation |
Auger, G, Martin, L, Bertrand, J, Ferrari, P, Fanchon, E, Vaganay, S, Pétillot, Y, van Heijenoort, J, Blanot, D and Dideberg, O (1998) Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli. Protein Expr. Purif. 13:23-9 |
|---|---|
| Abstract |
The UDP-N-acetylmuramoyl-L-alanine:D-glutamate ligase from Escherichia coli, an enzyme involved in the biosynthesis of the bacterial peptidoglycan monomer unit, was overproduced and purified to homogeneity on a large scale, yielding 4 mg of protein per liter of bacterial culture. Crystals of the complex with the substrate UDP-MurNAc-L-Ala were grown by the hanging drop method using ammonium sulfate as the precipitant. They are tetragonal with cell dimensions a = b = 65.5 A and c = 134.59 A, space group P4(1) or P4(3), and contain one monomer of 46,842 Da in the asymmetric unit. In order to use the multiple-wavelength anomalous diffraction method for phasing, a selenomethionine derivative of the protein has also been overproduced, purified, and crystallized. |
| Links |
PubMed Online version:10.1006/prep.1997.0850 |
| Keywords |
Amino Acid Sequence; Cloning, Molecular; Crystallography, X-Ray; Enzyme Stability; Escherichia coli/genetics; Mass Spectrometry; Molecular Sequence Data; Peptide Synthases/chemistry; Peptide Synthases/genetics; Peptide Synthases/isolation & purification; Recombinant Proteins/chemistry; Recombinant Proteins/genetics; Recombinant Proteins/isolation & purification; Selenomethionine/chemistry |
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