Purification, crystallization and preliminary X-ray analysis of Escherichia coli UDP-N-acetylmuramoyl:l-alanine ligase (MurC)

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 59; no. 8; pp. 1510 - 1513
• Main Authors: Deva, Taru, Pryor, KellyAnn D., Leiting, Barbara, Baker, Edward N., Smith, Clyde A.
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England Munksgaard International Publishers 01.08.2003
• Subjects: Adenosine Triphosphate - chemistry; amide-bond ligases; Carbohydrates - chemistry; Crystallization; Crystallography, X-Ray; Escherichia coli - enzymology; Escherichia coli - metabolism; Light; Peptide Synthases - chemistry; Peptide Synthases - isolation & purification; peptoglycan; Scattering, Radiation; UDP-N-acetylmuramoyl:l-alanine ligase; X-Ray Diffraction; X-Rays
• ISSN: 1399-0047; 0907-4449; 1399-0047
• DOI: 10.1107/S090744490301285X

UDP‐N‐acetylmuramoyl:l‐alanine ligase (MurC) is involved in the pathway leading from UDP‐N‐glucosamine to the UDP‐N‐acetylmuramoyl:pentapeptide unit, which is the building block for the peptidoglycan layer found in all bacterial cell walls. The pathways leading to the biosynthesis of the peptidoglycan layer are important targets for the development of novel antibiotics, since animal cells do not contain these pathways. MurC is the first of four similar ATP‐dependent amide‐bond ligases which share primary and tertiary structural similarities. The crystal structures of three of these have been determined by X‐ray crystallography, giving insights into the binding of the carbohydrate substrate and the ATP. Diffraction‐quality crystals of the enzyme MurC have been obtained in both native and selenomethionine forms and X‐ray diffraction data have been collected at the Se edge at a synchrotron source. The crystals are orthorhombic, with unit‐cell parameters a = 73.9, b = 93.6, c = 176.8 Å, and diffraction has been observed to 2.6 Å resolution.