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Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli.
Protein Expr Purif. 1998 Jun; 13(1):23-9.PE

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.

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Authors+Show Affiliations

Auger G
Unité de Recherche Associée 1131, Centre National de la Recherche Scientifique, Biochimie Moléculaire et Cellulaire, Université de Paris-Sud, Orsay, France.
Martin L
No affiliation info available
Bertrand J
No affiliation info available
Ferrari P
No affiliation info available
Fanchon E
No affiliation info available
Vaganay S
No affiliation info available
Pétillot Y
No affiliation info available
van Heijenoort J
No affiliation info available
Blanot D
No affiliation info available
Dideberg O
No affiliation info available

MeSH

Amino Acid SequenceCloning, MolecularCrystallography, X-RayEnzyme StabilityEscherichia coliMass SpectrometryMolecular Sequence DataPeptide SynthasesRecombinant ProteinsSelenomethionine

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

9631510

Citation

Auger, G, et al. "Large-scale Preparation, Purification, and Crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate Ligase From Escherichia Coli." Protein Expression and Purification, vol. 13, no. 1, 1998, pp. 23-9.
Auger G, Martin L, Bertrand J, et al. Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli. Protein Expr Purif. 1998;13(1):23-9.
Auger, G., Martin, L., Bertrand, J., Ferrari, P., Fanchon, E., Vaganay, S., Pétillot, Y., van Heijenoort, J., Blanot, D., & Dideberg, O. (1998). Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli. Protein Expression and Purification, 13(1), 23-9.
Auger G, et al. Large-scale Preparation, Purification, and Crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate Ligase From Escherichia Coli. Protein Expr Purif. 1998;13(1):23-9. PubMed PMID: 9631510.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Large-scale preparation, purification, and crystallization of UDP-N-acetylmuramoyl-L-alanine: D-glutamate ligase from Escherichia coli. AU - Auger,G, AU - Martin,L, AU - Bertrand,J, AU - Ferrari,P, AU - Fanchon,E, AU - Vaganay,S, AU - Pétillot,Y, AU - van Heijenoort,J, AU - Blanot,D, AU - Dideberg,O, PY - 1998/6/19/pubmed PY - 1998/6/19/medline PY - 1998/6/19/entrez SP - 23 EP - 9 JF - Protein expression and purification JO - Protein Expr Purif VL - 13 IS - 1 N2 - 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. SN - 1046-5928 UR - https://www.unboundmedicine.com/medline/citation/9631510/Large_scale_preparation_purification_and_crystallization_of_UDP_N_acetylmuramoyl_L_alanine:_D_glutamate_ligase_from_Escherichia_coli_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S1046-5928(97)90850-0 DB - PRIME DP - Unbound Medicine ER -