Allosteric Competitive Inhibitors of the Glucose-1-phosphate Thymidylyltransferase (RmlA) from Pseudomonas aeruginosa

Magnus Stephen Alphey, Lisa Pirrie, Leah S. Torrie, Wassila Abdelli Boulkeroua, Mary Gardiner, Aurijit Sarkar, Marko Maringer, Wulf Oehlmann, Ruth Brenk, Michael S. Scherman, Michael McNeil, Martin Rejzek, Robert Andrew Field, Mahavir Singh, David Gray, Nicholas James Westwood, Jim Naismith

Research output: Contribution to journalArticlepeer-review

22 Citations (Scopus)

Abstract

Glucose-1-phosphate thymidylyltransferase (RmlA) catalyzes the condensation of glucose-1-phosphate (G1P) with deoxy-thymidine triphosphate (dTTP) to yield dTDP-D-glucose and pyrophosphate. This is the first step in the L-rhamnose biosynthetic pathway. L-Rhamnose is an important component of the cell wall of many microorganisms, including Mycobacterium tuberculosis and Pseudomonas aeruginosa. Here we describe the first nanomolar inhibitors of P. aeruginosa RmlA. These thymine analogues were identified by high-throughput screening and subsequently optimized by a combination of protein crystallography, in silico screening, and synthetic chemistry. Some of the inhibitors show inhibitory activity against M. tuberculosis, The inhibitors do not bind at the active site of RmlA but bind at a second site active site. Despite this, the compounds act as competitive inhibitors of G1P but with high cooperativity. This novel behavior was probed by structural analysis, which suggests that the inhibitors work by preventing RmlA from undergoing the conformational change key to its ordered bi-bi mechanism.

Original languageEnglish
Pages (from-to)387-396
Number of pages10
JournalACS Chemical Biology
Volume8
Issue number2
Early online date8 Nov 2012
DOIs
Publication statusPublished - Feb 2013

Keywords

  • RECEPTOR
  • MYCOBACTERIUM-TUBERCULOSIS
  • NUCLEOTIDYLYLTRANSFERASE
  • CATALYTIC MECHANISM
  • DRUG DISCOVERY
  • RESISTANT
  • CRYSTAL-STRUCTURE
  • GLUCOSE PYROPHOSPHORYLASE
  • MODULATORS
  • RHAMNOSE SYNTHESIS

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