Cooperativity in Escherichia coli L-Threonine Dehydrogenase and Its Inhibition by an Antibacterial N-Pyridylpyrazolone Derivative

Antibiotic resistance is an increasing concern in modern healthcare. Therefore, it is important to identify novel antimicrobial agents and new molecular targets for such compounds. Here, we describe the identification of an N-pyridylpyrazolone derivative, 4-(2-aminoethyl)-2-(pyridin-2-yl)-1,2-dihydro-3H-pyrazol-3-one dihydrochloride (compound 1), which is effective against Gram-positive and Gram-negative bacteria and inhibits the enzymatic activity of Escherichia coli L-threonine dehydrogenase (TDH). To characterize its interaction with compound 1, TDH was overexpressed in E. coli. The recombinant enzyme was shown to exist in dilute solution in equilibrium between dimeric and tetrameric forms, with a Kd value for the dimer/tetramer transition of 3 ± 1 nM, and to bind L-threonine cooperatively with a Hill coefficient of 1.4. Compound 1 acted as a partial mixed inhibitor of TDH with an EC50 value of 47 ± 16 µM and did not affect the equilibrium between oligomeric states. Altogether, these findings identify compound 1 as a promising starting point for the development of novel antibiotics and as a tool compound for studying the functional properties of TDH.

• Publication year

  2025

• Venue

  International Journal of Molecular Sciences

• Publication date

  2025-12-01

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.3390/ijms262311751PMID 41373896PMCID 12693531
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

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• New look at hemoglobin allostery.

  2015influential reference
• Origin and evolution of medium chain alcohol dehydrogenases.

  2013cited by this paper
• One-pot parallel solution-phase synthesis of 1-substituted 4-(2-aminoethyl)-1H-pyrazol-5-ols.

  2008cited by this paper
• A simple synthesis of 4-(2-aminoethyl)-5-hydroxy-1H-pyrazoles

  2007influential reference
• Identification of C-terminal motifs responsible for transmission of inhibition by ATP of mammalian phosphofructokinase, and their contribution to other allosteric effects.

  2004influential reference
• The human L-threonine 3-dehydrogenase gene is an expressed pseudogene

  2002cited by this paper
• Kinetic Cooperativity of Human Liver Alcohol Dehydrogenase γ2 *

  2000cited by this paper
• Kinetic cooperativity of human liver alcohol dehydrogenase gamma(2).

  2000cited by this paper
• Investigation of a catalytic zinc binding site in Escherichia coli L-threonine dehydrogenase by site-directed mutagenesis of cysteine-38.

  1998cited by this paper
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  1998cited by this paper
• Functional analysis of E. coli threonine dehydrogenase by means of mutant isolation and characterization.

  1995cited by this paper
• A super-family of medium-chain dehydrogenases/reductases (MDR). Sub-lines including zeta-crystallin, alcohol and polyol dehydrogenases, quinone oxidoreductase enoyl reductases, VAT-1 and other proteins.

  1994cited by this paper
• Threonine formation via the coupled activity of 2-amino-3-ketobutyrate coenzyme A lyase and threonine dehydrogenase

  1993cited by this paper
• L-threonine dehydrogenase from Escherichia coli. Identification of an active site cysteine residue and metal ion studies.

  1991cited by this paper
• L-threonine dehydrogenase from Escherichia coli K-12: thiol-dependent activation by Mn2+.

  1986cited by this paper
• L-threonine dehydrogenase. Purification and properties of the homogeneous enzyme from Escherichia coli K-12.

  1981cited by this paper
• The allosteric regulation of hexokinase C from amphibian liver.

  1976influential reference
• ON THE NATURE OF ALLOSTERIC TRANSITIONS: A PLAUSIBLE MODEL.

  1965influential reference

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