scholarly journals MgF3− and AlF4− transition state analogue complexes of yeast phosphoglycerate kinase

2017 ◽  
Vol 95 (2) ◽  
pp. 295-303
Author(s):  
Nicole E. McCormick ◽  
Stephanie M. Forget ◽  
Raymond T. Syvitski ◽  
David L. Jakeman
Keyword(s):  
Nmr Spectroscopy ◽  
Transition State ◽  
Chemical Shifts ◽  
Phosphoglycerate Kinase ◽  
Indirect Evidence ◽  
Metal Fluoride ◽  
Binding Affinities ◽  
Transition State Analogue ◽  
Transition State Analogues ◽  
Proton Chemical Shifts

The phospho-transfer mechanism of yeast phosphoglycerate kinase (PGK) has been probed through formation of trifluoromagnesate (MgF3−) and tetrafluoroaluminate (AlF4−) transition state analogue complexes and analyzed using 19F, 1H waterLOGSY and 1H chemical shift perturbation NMR spectroscopy. We observed the first 19F NMR spectroscopic evidence for the formation of metal fluoride transition state analogues of yeast PGK and also observed significant changes to proton chemical shifts of PGK in the presence, but not in the absence, of fluoride upon titration of ligands, providing indirect evidence of the formation of a closed ternary transition state. WaterLOGSY NMR spectroscopy experiments using an uncompetitive model were used in an attempt to measure ligand binding affinities within the transition state analogue complexes.

scholarly journals Inhibition of Clostridium difficile TcdA and TcdB toxins with transition state analogues

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Ashleigh S. Paparella ◽  
Briana L. Aboulache ◽  
Rajesh K. Harijan ◽  
Kathryn S. Potts ◽  
Peter C. Tyler ◽  
...  
Keyword(s):  
Clostridium Difficile ◽  
Transition State ◽  
Rho Gtpases ◽  
Mammalian Cells ◽  
Bacterial Infections ◽  
The United States ◽  
Transition State Analogue ◽  
Transition State Analogues ◽  
Transition State Analogue Inhibitors ◽  
Catalyzed Reactions

AbstractClostridium difficile causes life-threatening diarrhea and is the leading cause of healthcare-associated bacterial infections in the United States. TcdA and TcdB bacterial toxins are primary determinants of disease pathogenesis and are attractive therapeutic targets. TcdA and TcdB contain domains that use UDP-glucose to glucosylate and inactivate host Rho GTPases, resulting in cytoskeletal changes causing cell rounding and loss of intestinal integrity. Transition state analysis revealed glucocationic character for the TcdA and TcdB transition states. We identified transition state analogue inhibitors and characterized them by kinetic, thermodynamic and structural analysis. Iminosugars, isofagomine and noeuromycin mimic the transition state and inhibit both TcdA and TcdB by forming ternary complexes with Tcd and UDP, a product of the TcdA- and TcdB-catalyzed reactions. Both iminosugars prevent TcdA- and TcdB-induced cytotoxicity in cultured mammalian cells by preventing glucosylation of Rho GTPases. Iminosugar transition state analogues of the Tcd toxins show potential as therapeutics for C. difficile pathology.

Transition State Analogue Structures of Human Phosphoglycerate Kinase Establish the Importance of Charge Balance in Catalysis

2010 ◽  
Vol 132 (18) ◽  
pp. 6507-6516 ◽  
Author(s):  
Matthew J. Cliff ◽  
Matthew W. Bowler ◽  
Andrea Varga ◽  
James P. Marston ◽  
Judit Szabó ◽  
...  
Keyword(s):  
Transition State ◽  
Phosphoglycerate Kinase ◽  
Charge Balance ◽  
Transition State Analogue

The acidity of β-phosphoglucomutase monofluoromethylenephosphonate ligands probed by NMR spectroscopy and quantum mechanical methods

2016 ◽  
Vol 94 (11) ◽  
pp. 902-908 ◽  
Author(s):  
Stephanie M. Forget ◽  
Eric A.C. Bushnell ◽  
Russell J. Boyd ◽  
David L. Jakeman
Keyword(s):  
Nmr Spectroscopy ◽  
Transition State ◽  
Quantum Mechanical ◽  
Phosphoryl Transfer ◽  
Quantum Mechanical Calculations ◽  
Transition State Analogues ◽  
Mechanical Methods ◽  
Insight Into ◽  
Computational Quantum

We recently described the binding of 1-β-phosphonomethylene-1-deoxy-d-glucopyranose, (S)-1-β-phosphonofluoromethylene-1-deoxy-D-glucopyranose (βG1CFSP), and (R)-1-β-phosphonofluoromethylene-1-deoxy-d-glucopyranose (βG1CFRP) to the enzyme β-phosphoglucomutase as transition state analogues of phosphoryl transfer through formation of stable MgF3− and AlF4− complexes (Proc. Natl. Acad. Sci. U.S.A. 2014, 111, 12384). Herein, we describe in detail the five-fold difference in acidity (pKa2) for the (S)- and (R)-configured diastereomeric fluorophosphonates through a series of NMR spectroscopy experiments. The differences in acidity were corroborated using computational quantum mechanical calculations to determine structures of lowest energy conformers and provide insight into why the (S) isomer is substantially more acidic.

scholarly journals Observing enzyme ternary transition state analogue complexes by 19F NMR spectroscopy

2017 ◽  
Vol 8 (12) ◽  
pp. 8427-8434 ◽  
Author(s):  
Anna Ampaw ◽  
Madison Carroll ◽  
Jill von Velsen ◽  
Debabrata Bhattasali ◽  
Alejandro Cohen ◽  
...  
Keyword(s):  
Nmr Spectroscopy ◽  
Transition State ◽  
19F Nmr ◽  
19F Nmr Spectroscopy ◽  
Transition State Analogue ◽  
Catalytically Active

Ternary transition state analogue (TSA) complexes probing the isomerization of β-d-glucose 1-phosphate (G1P) into d-glucose 6-phosphate (G6P) catalyzed by catalytically active, fluorinated (5-fluorotryptophan), β-phosphoglucomutase (βPGM) have been observed directly by 19F NMR spectroscopy.

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