title: Kinetic evaluation of glucose 1-phosphate analogues with a thymidylyltransferase using a continuous coupled enzyme assay.
abstract: Cps2L, a thymidylytransferase, is the first enzyme in Streptococcus pneumoniae L-rhamnose biosynthesis and an antibacterial target. We herein report the evaluation of six sugar phosphate analogues selected to further probe Cps2L substrate tolerance. A modified continuous spectrophotometric assay was employed for facile detection of pyrophosphate (PPi) released from nucleotidylyltransfase-catalysed condensation of sugar 1-phosphates and nucleoside triphosphates to produce sugar nucleotides. Additionally, experiments using waterLOGSY NMR spectroscopy were investigated as a complimentary method to evaluate binding affinity to Cps2L.
authors.affiliation: Department of Chemistry, Dalhousie University, 6274 Coburg Rd, PO Box 15, 000, Halifax, Nova Scotia B3H 4R2, Canada. David.Jakeman@dal.ca.
authors.forename: S M
authors.initials: SM
authors.lastname: Forget
authors.forename: A
authors.initials: A
authors.lastname: Jee
authors.forename: D A
authors.initials: DA
authors.lastname: Smithen
authors.forename: R
authors.initials: R
authors.lastname: Jagdhane
authors.forename: S
authors.initials: S
authors.lastname: Anjum
authors.forename: S A
authors.initials: SA
authors.lastname: Beaton
authors.forename: D R J
authors.initials: DR
authors.lastname: Palmer
authors.forename: R T
authors.initials: RT
authors.lastname: Syvitski
authors.forename: D L
authors.initials: DL
authors.lastname: Jakeman
mesh_terms: D000900:Anti-Bacterial Agents / Q000138:chemical synthesis / Q000737:chemistry*; D001426:Bacterial Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry*; D011756:Diphosphates / Q000032:analysis; D057075:Enzyme Assays; D004791:Enzyme Inhibitors / Q000138:chemical synthesis / Q000737:chemistry*; D005958:Glucosephosphates / Q000737:chemistry*; D007700:Kinetics; D009713:Nucleotidyltransferases / Q000037:antagonists & inhibitors / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D013053:Spectrophotometry; D013296:Streptococcus pneumoniae / Q000737:chemistry / Q000201:enzymology
chemical_list: D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D011756:Diphosphates; D004791:Enzyme Inhibitors; D005958:Glucosephosphates; D011994:Recombinant Proteins; C107241:diphosphoric acid; C031590:glucose-1-phosphate; D009713:Nucleotidyltransferases; C020587:glucose-1-phosphate thymidylyltransferase