title: In vitro reconstruction of the biosynthetic pathway of peptidoglycan cytoplasmic precursor in Pseudomonas aeruginosa.
abstract: Bacterial peptidoglycan is the cell wall component responsible for maintaining cell integrity against osmotic pressure. Biosynthesis of the cytoplasmic precursor UDP-N-acetylmuramyl pentapeptide is catalyzed by the Mur enzymes. Genomic analysis of the three regions encoding Mur proteins was achieved. We have cloned and over-expressed the murA, -B, -D, -E and -F genes of Pseudomonas aeruginosa in pET expression system by adding a His-Tag to the C-termini of the proteins. Mur proteins were purified to homogeneity by a single chromatographic step on affinity nickel columns. Protein identities were verified through N-terminal sequencing. Enzyme activity was proved by the identification of the pathway's final product.
authors.affiliation: Centre de Recherche sur la Fonction, Structure et Ingénierie des Protéines, Faculté de Médecine, Ste-Foy, QC, Canada.
authors.forename: A
authors.initials: A
authors.lastname: El Zoeiby
authors.forename: F
authors.initials: F
authors.lastname: Sanschagrin
authors.forename: P C
authors.initials: PC
authors.lastname: Havugimana
authors.forename: A
authors.initials: A
authors.lastname: Garnier
authors.forename: R C
authors.initials: RC
authors.lastname: Levesque
mesh_terms: D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D002473:Cell Wall / Q000378:metabolism; D002846:Chromatography, Affinity; D003001:Cloning, Molecular; D005798:Genes, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000096:biosynthesis* / Q000235:genetics; D011550:Pseudomonas aeruginosa / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein
chemical_list: D001426:Bacterial Proteins; D010457:Peptidoglycan; D011993:Recombinant Fusion Proteins