Functional and structural basis of E. coli enolase inhibition by SF2312: a mimic of the carbanion intermediate

  • Sci Rep. 2019 Nov 19;9(1):17106. doi: 10.1038/s41598-019-53301-3.
Jolanta Krucinska  1 ,  Michael N Lombardo  1 ,  Heidi Erlandsen  2 ,  Akram Hazeen  3 ,  Searle S Duay  3 ,  Jason G Pattis  4 ,  Victoria L Robinson  4 ,  Eric R May  4 ,  Dennis L Wright  5  6
Affiliations
  • 1. Department of Pharmaceutical Sciences, University of Connecticut, 69 North Eagleville Road, Storrs, Connecticut, 06269, United States.
  • 2. Center for Open Research Resources & Equipment (COR2E), University of Connecticut, 91 North Eagleville Road, Storrs, Connecticut, 06269, United States.
  • 3. Department of Chemistry, University of Connecticut, 55 North Eagleville Road, Storrs, Connecticut, 06269, United States.
  • 4. Department of Molecular and Cellular Biology, University of Connecticut, 91 North Eagleville Road, Storrs, Connecticut, 06269, United States.
  • 5. Department of Pharmaceutical Sciences, University of Connecticut, 69 North Eagleville Road, Storrs, Connecticut, 06269, United States. [email protected].
  • 6. Department of Chemistry, University of Connecticut, 55 North Eagleville Road, Storrs, Connecticut, 06269, United States. [email protected].
PMID: 31745118 DOI: 10.1038/s41598-019-53301-3
Abstract

Many years ago, the natural secondary metabolite SF2312, produced by the actinomycete Micromonospora, was reported to display broad spectrum Antibacterial properties against both Gram-positive and Gram-negative bacteria. Recent studies have revealed that SF2312, a natural phosphonic acid, functions as a potent inhibitor of human Enolase. The mechanism of SF2312 inhibition of Bacterial enolase and its role in Bacterial growth and reproduction, however, have remained elusive. In this work, we detail a structural analysis of E. coli Enolase bound to both SF2312 and its oxidized imide-form. Our studies support a model in which SF2312 acts as an analog of a high energy intermediate formed during the catalytic process. Biochemical, biophysical, computational and kinetic characterization of these compounds confirm that altering features characteristic of a putative carbanion (enolate) intermediate significantly reduces the potency of enzyme inhibition. When SF2312 is combined with fosfomycin in the presence of glucose-6 phosphate, significant synergy is observed. This suggests the two agents could be used as a potent combination, targeting distinct cellular mechanism for the treatment of Bacterial infections. Together, our studies rationalize the structure-activity relationships for these phosphonates and validate Enolase as a promising target for Antibiotic discovery.

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