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Cited by 20Biochemical and Molecular ResearchMicrobial Natural Products and BiosynthesisChemical Reaction Mechanisms

An Open and Shut Case: The Interaction of Magnesium with MST Enzymes

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Meneely, Kathleen M., Sundlov, Jesse A., Gulick, Andrew M., Moran, Graham R., Lamb, Audrey L.·2016·Journal of the American Chemical Society
DOI 10.1021/jacs.6b05134PMC PMC5029964
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Abstract

The shikimate pathway of bacteria, fungi, and plants generates chorismate, which is drawn into biosynthetic pathways that form aromatic amino acids and other important metabolites, including folates, menaquinone, and siderophores. Many of the pathways initiated at this branch point transform chorismate using an MST enzyme. The MST enzymes (menaquinone, siderophore, and tryptophan biosynthetic enzymes) are structurally homologous and magnesium-dependent, and all perform similar chemical permutations to chorismate by nucleophilic addition (hydroxyl or amine) at the 2-position of the ring, inducing displacement of the 4-hydroxyl. The isomerase enzymes release isochorismate or aminodeoxychorismate as the product, while the synthase enzymes also have lyase activity that displaces pyruvate to form either salicylate or anthranilate. This has led to the hypothesis that the isomerase and lyase activities performed by the MST enzymes are functionally conserved. Here we have developed tailored pre-steady-state approaches to establish the kinetic mechanisms of the isochorismate and salicylate synthase enzymes of siderophore biosynthesis. Our data are centered on the role of magnesium ions, which inhibit the isochorismate synthase enzymes but not the salicylate synthase enzymes. Prior structural data have suggested that binding of the metal ion occludes access or egress of substrates. Our kinetic data indicate that for the production of isochorismate, a high magnesium ion concentration suppresses the rate of release of product, accounting for the observed inhibition and establishing the basis of the ordered-addition kinetic mechanism. Moreover, we show that isochorismate is channeled through the synthase reaction as an intermediate that is retained in the active site by the magnesium ion. Indeed, the lyase-active enzyme has 3 orders of magnitude higher affinity for the isochorismate complex relative to the chorismate complex. Apparent negative-feedback inhibition by ferrous ions is documented at nanomolar concentrations, which is a potentially physiologically relevant mode of regulation for siderophore biosynthesis in vivo.

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  • This Mg2+ can be readily displaced by Fe2+ or Mn2+ leading to enzyme inhibition, suggesting that this Mg2+ is labile.

    “This Mg2+ can be readily displaced by Fe2+ or Mn2+ leading to enzyme inhibition,81,82 suggesting that this Mg2+ is labile.”

    From Metals in Motion: Understanding Labile Metal Pools in Bacteria (2025) · cites this paper as [81]

    The Results report that Fe(II) binds at the catalytic magnesium site and inhibits MST enzymes in the presence of magnesium. The Discussion says magnesium dissociates after each catalytic cycle, consistent with its being labile. The cited passages do not establish displacement or inhibition by Mn(II), or show that Fe(II) directly displaces magnesium already bound to the enzyme. (LLM, conf=0.87)

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  • Enzymes of the menaquinone, siderophore, and tryptophan (MST) family bind Mg2+ reversibly with a Kd of ~0.2 mM.

    “Similarly, enzymes of the menaquinone, siderophore, and tryptophan (MST) family bind Mg2+ reversibly with a Kd of ∼0.2 mM.81”

    From Metals in Motion: Understanding Labile Metal Pools in Bacteria (2025) · cites this paper as [81]

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Authors
Meneely, K. M., Sundlov, J. A., Gulick, A. M., Moran, G. R., & Lamb, A. L.
Affiliations
  • University of Kansas (US)
  • Hauptman-Woodward Medical Research Institute (US)
  • University of Wisconsin–Milwaukee (US)
Corresponding author
  • Moran, Graham R.
  • Lamb, Audrey L.
ORCID
Venue
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Published
2016-07-03
Volume
138
Issue
29
Pages
9277-9293
ISSN
  • 0002-7863 (ISSN-L)
  • 1520-5126
DOI
10.1021/jacs.6b05134
PMC
PMC5029964
OpenAlex
W2463282447
Cited by
20
References
1
Citations by year
2026: 2 · 2025: 1 · 2024: 2 · 2023: 2 · 2022: 3 · 2021: 2 · 2020: 4 · 2018: 3 · 2017: 1
FWCI
0.57
Type
journal article
Language
en
Open access
Licence
Locations
  • Journal of the American Chemical Society (publishedVersion, other-oa)
  • PubMed Central (publishedVersion)
  • PubMed (publishedVersion)
Funders
  • National Institute of General Medical Sciences (R01 GM116957)
  • National Institute of Allergy and Infectious Diseases (K02 AI093675, R01 AI77725)
  • National Center for Research Resources (P20 RR016475)
  • Division of Chemistry (CHE 1402475, CHE 1403293)
  • University of Wisconsin-Milwaukee (NA)
  • General Research Fund, University of Kansas (230189)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Biochemical and Molecular Research
  • Health Sciences › Medicine › Pharmacology › Microbial Natural Products and Biosynthesis
  • Physical Sciences › Chemistry › Organic Chemistry › Chemical Reaction Mechanisms
Keywords
shikimate pathway, chorismate, menaquinone biosynthesis, siderophore biosynthesis, tryptophan biosynthesis, isochorismate synthase, enzyme kinetics, active site metal ions, aromatic amino acid biosynthesis, pre-steady-state kinetics
MeSH terms
Binding Sites, Chorismic Acid, Kinetics, Magnesium, Models, Molecular, Protein Binding, Tryptophan, Siderophores, Intramolecular Transferases, Catalytic Domain, Vitamin K 2

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