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

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.

Authors:
 [1];  [1];  [2];  [3];  [4];  [5];  [5];  [5];  [6]
  1. Univ. of Connecticut, Storrs, CT (United States). Dept. of Pharmaceutical Sciences
  2. Univ. of Connecticut, Storrs, CT (United States). Center for Open Research Resources & Equipment (COR2E)
  3. Univ. of Connecticut, Storrs, CT (United States). Dept. of Chemistry
  4. Univ. of Connecticut, Storrs, CT (United States). Dept. of Chemistry
  5. Univ. of Connecticut, Storrs, CT (United States). Dept. of Molecular and Cellular Biology
  6. Univ. of Connecticut, Storrs, CT (United States). Dept. of Pharmaceutical Sciences; Univ. of Connecticut, Storrs, CT (United States). Dept. of Chemistry
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
OSTI Identifier:
1624507
Grant/Contract Number:  
AC02-76SF00515; 1R21AI140734-01; R35-GM119762; P41GM103393
Resource Type:
Accepted Manuscript
Journal Name:
Scientific Reports
Additional Journal Information:
Journal Volume: 9; Journal Issue: 1; Journal ID: ISSN 2045-2322
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; Science & Technology - Other Topics

Citation Formats

Krucinska, Jolanta, Lombardo, Michael N., Erlandsen, Heidi, Hazeen, Akram, Duay, Searle S., Pattis, Jason G., Robinson, Victoria L., May, Eric R., and Wright, Dennis L. Functional and structural basis of E. coli enolase inhibition by SF2312: a mimic of the carbanion intermediate. United States: N. p., 2019. Web. https://doi.org/10.1038/s41598-019-53301-3.
Krucinska, Jolanta, Lombardo, Michael N., Erlandsen, Heidi, Hazeen, Akram, Duay, Searle S., Pattis, Jason G., Robinson, Victoria L., May, Eric R., & Wright, Dennis L. Functional and structural basis of E. coli enolase inhibition by SF2312: a mimic of the carbanion intermediate. United States. https://doi.org/10.1038/s41598-019-53301-3
Krucinska, Jolanta, Lombardo, Michael N., Erlandsen, Heidi, Hazeen, Akram, Duay, Searle S., Pattis, Jason G., Robinson, Victoria L., May, Eric R., and Wright, Dennis L. Tue . "Functional and structural basis of E. coli enolase inhibition by SF2312: a mimic of the carbanion intermediate". United States. https://doi.org/10.1038/s41598-019-53301-3. https://www.osti.gov/servlets/purl/1624507.
@article{osti_1624507,
title = {Functional and structural basis of E. coli enolase inhibition by SF2312: a mimic of the carbanion intermediate},
author = {Krucinska, Jolanta and Lombardo, Michael N. and Erlandsen, Heidi and Hazeen, Akram and Duay, Searle S. and Pattis, Jason G. and Robinson, Victoria L. and May, Eric R. and Wright, Dennis L.},
abstractNote = {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.},
doi = {10.1038/s41598-019-53301-3},
journal = {Scientific Reports},
number = 1,
volume = 9,
place = {United States},
year = {2019},
month = {11}
}

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