Kinetic Modelling of GlmU Reactions – Prioritization of Reaction for Therapeutic Application

  title={Kinetic Modelling of GlmU Reactions – Prioritization of Reaction for Therapeutic Application},
  author={Vivek Kumar Singh and Kaveri S Das and Kothandaraman Seshadri},
  journal={PLoS ONE},
Mycobacterium tuberculosis(Mtu), a successful pathogen, has developed resistance against the existing anti-tubercular drugs necessitating discovery of drugs with novel action. Enzymes involved in peptidoglycan biosynthesis are attractive targets for antibacterial drug discovery. The bifunctional enzyme mycobacterial GlmU (Glucosamine 1-phosphate N-acetyltransferase/ N-acetylglucosamine-1-phosphate uridyltransferase) has been a target enzyme for drug discovery. Its C- and N- terminal domains… 
Inhibition studies on Mycobacterium tuberculosis N-acetylglucosamine-1-phosphate uridyltransferase (GlmU).
The most potent inhibitor in this series exhibited an IC50 of 74 μM against GlmU uridyltransferase activity and serves as a promising starting point for the discovery of more potent inhibitors.
Mutational analysis to identify the residues essential for the acetyltransferase activity of GlmU in Bacillus subtilis
The novel and attractive antimicrobial drug target, glucosamine-1-phosphate acetyltransferase/N-acetylglucosamine-1-phosphate uridyltransferase (GlmU), is a bifunctional enzyme that catalyzes two
Purification and biochemical characterisation of GlmU from Yersinia pestis
A biochemical characterisation of the Yersinia GlmU showed the acetyltransferase activity, which is strongly increased in the presence of reducing agent, was shown to be susceptible to oxidation and thiol-specific reagents.
Depletion of M. tuberculosis GlmU from Infected Murine Lungs Effects the Clearance of the Pathogen
It is found that absence of GlmUMtb leads to extensive perturbation of bacterial morphology and substantial reduction in cell wall thickness under normoxic as well as hypoxic conditions, which establishes Glm UMtb as a strong candidate for intervention measures against established tuberculosis infections.
Structure-based design of diverse inhibitors of Mycobacterium tuberculosis N-acetylglucosamine-1-phosphate uridyltransferase: combined molecular docking, dynamic simulation, and biological activity
Design and screening of nine inhibitors against UTP and NAcGlc-1-P of uridyltransferase active site of glmUMtb are reported, which can serve as a starting point in the drug discovery processes against Mycobacterium tuberculosis.
GlmU from Mycobacterium tuberculosis – Structure, Function, and the Role of Metal Ions in Catalysis
N-Acetylglucosamine-1-phosphate uridyltransferase (GlmU) forms a biological trimer. Each monomer has two independent domains that catalyze two independent enzymatic reactions. The C-terminal domain
Exploring MDR-TB inhibitory potential of 4-amino quinazolines as Mycobacterium tuberculosis N -acetylglucosamine-1-phosphate uridyltransferase (GlmU MTB) inhibitors.
The 4-amino quinazolines as M. tuberculosis N -acetylglucosamine-1-phosphate uridyltransferase (GlmU MTB) inhibitors to overcome the problem of the MDR-TB are reported.
A simple and rapid method for measuring α-D-phosphohexomutases activity by using anion-exchange chromatography coupled with an electrochemical detector
HPAEC-PAD is a more rapid and simple method than the traditional coupled assays given its high specificity and sensitivity, and will certainly bring convenience to further research of α-D-phosphohexomutases superfamily.


Structure and function of GlmU from Mycobacterium tuberculosis.
Enzyme activities were characterized by (1)H NMR and suggest that the presence of acetyl-coenzyme A has an inhibitory effect on uridyltransferase activity.
High-Throughput Screening Identifies Novel Inhibitors of the Acetyltransferase Activity of Escherichia coli GlmU
An absorbance-based assay is developed to screen diverse chemical libraries in high throughput for inhibitors to the acetyltransferase reaction of Escherichia coli GlmU and outlines the utility of catalytic variants in targeting specific activities of bifunctional enzymes in high-throughput screens.
Kinetic modeling of tricarboxylic acid cycle and glyoxylate bypass in Mycobacterium tuberculosis, and its application to assessment of drug targets
  • V. Singh, I. Ghosh
  • Biology, Medicine
    Theoretical Biology and Medical Modelling
  • 2006
BackgroundTargeting persistent tubercule bacilli has become an important challenge in the development of anti-tuberculous drugs. As the glyoxylate bypass is essential for persistent bacilli,
Kinetic properties of Mycobacterium tuberculosis bifunctional GlmU
Two accurate and simple colorimetric assays based on 96-well microtiter plate were developed to measure the kinetic properties of bifunctional GlmU including initial velocity, optimal temperature, optimal pH, the effect of Mg2+, and the kinetic parameters that will facilitate high-throughput screening of Glm U inhibitors.
Structure of a small‐molecule inhibitor complexed with GlmU from Haemophilus influenzae reveals an allosteric binding site
Analysis of the mechanistic model of the uridyltransferase reaction suggests that the binding of this allosteric inhibitor prevents structural rearrangements that are required for the enzymatic reaction, thus providing a basis for structure‐guided design of a new class of mechanism‐based inhibitors of GlmU.
Prospects for Antiparasitic Drugs
Only uncompetitive inhibition of pyruvate export proves effective in the model; in all other cases studied, the effects on metabolite concentrations areLittle prospect of killing trypanosomes by depressing their glycolysis to a level incapable of sustaining life.
Expression, essentiality, and a microtiter plate assay for mycobacterial GlmU, the bifunctional glucosamine-1-phosphate acetyltransferase and N-acetylglucosamine-1-phosphate uridyltransferase.
Enzymatic assays showed that M. tuberculosis GlmU protein exhibits both glucosamine-1-phosphate acetyltransferase and N-acetylglucosamine -1- phosphate uridylyltransferase activities, which support the further development of M.culosis Glm U enzyme as a target for new anti-tuberculosis drugs.
Design and synthesis of novel cell wall inhibitors of Mycobacterium tuberculosis GlmM and GlmU.
M and GlmU are key enzymes in the biosynthesis of UDP-N-acetyl-d-glucosamine, an essential precursor of peptidoglycan and the rhamnose-GlcNAc linker region in the mycobacterial cell wall, and two analogs of GlcN-6-P and glucosamine-1-phosphate are synthesized as inhibitors of these enzymes.
Acetyltransfer precedes uridylyltransfer in the formation of UDP-N-acetylglucosamine in separable active sites of the bifunctional GlmU protein of Escherichia coli.
Kinetic studies demonstrated that a pre-steady-state lag in the production of UDP-N-acetylglucosamine from acetyl-CoA, UTP, and glucosamine-1-P was due to the release and accumulation of steady-state levels of the intermediate N-acetelglucOSamine- 1-P.
Genes required for mycobacterial growth defined by high density mutagenesis
The use of transposon site hybridization (TraSH) is described to comprehensively identify the genes required by the causative agent, Mycobacterium tuberculosis, for optimal growth, suggesting that the minimal gene set required for survival varies greatly between organisms with different evolutionary histories.