New Delhi Metallo‐Beta‐Lactamase Variants NDM‐4 and NDM‐12 from E. coli Clinical Isolates Exhibit Increased Activity and Stability

Williams, Cameron; VanPelt, Jamie; Poth, Robert; Cottingim, Kelsey; Stewart, Alesha; Nix, Jay C.; Fast, Walter; Tierney, David L.; Crowder, Michael W.; Page, Richard C.

doi:10.1096/fasebj.31.1_supplement.777.21

Title: New Delhi Metallo‐Beta‐Lactamase Variants NDM‐4 and NDM‐12 from E. coli Clinical Isolates Exhibit Increased Activity and Stability

Full Record
Other Related Research

Abstract

Metallo‐Beta‐Lactamases (MBLs) confer resistance to carbapenems, cephalosporins, and penicillins in several clinically relevant Gram‐negative bacteria including Acinetobacter, Pseudomonas aeruginosa, and many Enterobacteriaceae. New Delhi Metallo‐Beta‐Lactamases (NDMs) are amongst some of the most worrisome and prevalent MBLs. This family of MBLs is capable of hydrolyzing all generations of bicyclic beta‐lactams. Two new NDM variants, NDM‐4 and ‐12, have recently been identified in E. coli clinical isolates. NDM‐4 contains an M154L substitution while NDM‐12 bears both M154L and G222D mutations. Considering the clinical significance of Gram‐negative species possessing these enzymes, there is a need to characterize the structures and activities of these variants in order to provide a better basis for drug discovery efforts. We have found that NDM‐4 and NDM‐12 possess higher hydrolytic activity compared to NDM‐1. Our differential scanning fluorimetry studies suggest that the M154L mutation confers increased thermal stability and improved affinity for active site catalytic Zn metal ions. The additional G222D mutation of NDM‐12 further improves stability and Zn affinity. M154L, found in the secondary coordination sphere, accounts for changes in stability by acting as a buttress to the loop containing His ₁₂₂ , a direct Zn coordinating residue. G222D, on the other hand, is a mutation locatedmore »« less

Authors:

Williams, Cameron ^[1]; VanPelt, Jamie ^[1]; Poth, Robert ^[1]; Cottingim, Kelsey ^[2]; Stewart, Alesha ^[3]; Nix, Jay C. ^[4]; Fast, Walter ^[3]; Tierney, David L. ^[1]; Crowder, Michael W. ^[1]; Page, Richard C. ^[2]

Department of Chemistry and Biochemistry Miami University Oxford OH
Miami University Oxford OH
Medicinal Chemistry Division University of Texas at Austin Austin TX
Molecular Biology Consortium, Beamline 4.2.2 Lawrence Berkeley National Laboratory Berkeley CA

Publication Date:: Wed Oct 03 00:00:00 EDT 2018

Sponsoring Org.:: USDOE

OSTI Identifier:: 1787056

Grant/Contract Number:: DE‐AC03‐76SF00098

Resource Type:: Publisher's Accepted Manuscript

Journal Name:: FASEB Journal

Additional Journal Information:: Journal Name: FASEB Journal Journal Volume: 31 Journal Issue: S1; Journal ID: ISSN 0892-6638

Publisher:: FASEB

Country of Publication:: United States

Language:: English

Citation Formats


                    Williams, Cameron, VanPelt, Jamie, Poth, Robert, Cottingim, Kelsey, Stewart, Alesha, Nix, Jay C., Fast, Walter, Tierney, David L., Crowder, Michael W., and Page, Richard C. New Delhi Metallo‐Beta‐Lactamase Variants NDM‐4 and NDM‐12 from E. coli Clinical Isolates Exhibit Increased Activity and Stability.  United States: N. p., 2018. 
Web.  doi:10.1096/fasebj.31.1_supplement.777.21.

Copy to clipboard


                    Williams, Cameron, VanPelt, Jamie, Poth, Robert, Cottingim, Kelsey, Stewart, Alesha, Nix, Jay C., Fast, Walter, Tierney, David L., Crowder, Michael W., & Page, Richard C. New Delhi Metallo‐Beta‐Lactamase Variants NDM‐4 and NDM‐12 from E. coli Clinical Isolates Exhibit Increased Activity and Stability.  United States.  https://doi.org/10.1096/fasebj.31.1_supplement.777.21

Copy to clipboard


                    Williams, Cameron, VanPelt, Jamie, Poth, Robert, Cottingim, Kelsey, Stewart, Alesha, Nix, Jay C., Fast, Walter, Tierney, David L., Crowder, Michael W., and Page, Richard C. Wed .  
"New Delhi Metallo‐Beta‐Lactamase Variants NDM‐4 and NDM‐12 from E. coli Clinical Isolates Exhibit Increased Activity and Stability".  United States.  https://doi.org/10.1096/fasebj.31.1_supplement.777.21.

Copy to clipboard


                    
@article{osti_1787056,

  title        = {New Delhi Metallo‐Beta‐Lactamase Variants NDM‐4 and NDM‐12 from E. coli Clinical Isolates Exhibit Increased Activity and Stability},

  author       = {Williams, Cameron and VanPelt, Jamie and Poth, Robert and Cottingim, Kelsey and Stewart, Alesha and Nix, Jay C. and Fast, Walter and Tierney, David L. and Crowder, Michael W. and Page, Richard C.},

  abstractNote = {Metallo‐Beta‐Lactamases (MBLs) confer resistance to carbapenems, cephalosporins, and penicillins in several clinically relevant Gram‐negative bacteria including Acinetobacter, Pseudomonas aeruginosa, and many Enterobacteriaceae. New Delhi Metallo‐Beta‐Lactamases (NDMs) are amongst some of the most worrisome and prevalent MBLs. This family of MBLs is capable of hydrolyzing all generations of bicyclic beta‐lactams. Two new NDM variants, NDM‐4 and ‐12, have recently been identified in E. coli clinical isolates. NDM‐4 contains an M154L substitution while NDM‐12 bears both M154L and G222D mutations. Considering the clinical significance of Gram‐negative species possessing these enzymes, there is a need to characterize the structures and activities of these variants in order to provide a better basis for drug discovery efforts. We have found that NDM‐4 and NDM‐12 possess higher hydrolytic activity compared to NDM‐1. Our differential scanning fluorimetry studies suggest that the M154L mutation confers increased thermal stability and improved affinity for active site catalytic Zn metal ions. The additional G222D mutation of NDM‐12 further improves stability and Zn affinity. M154L, found in the secondary coordination sphere, accounts for changes in stability by acting as a buttress to the loop containing His 122 , a direct Zn coordinating residue. G222D, on the other hand, is a mutation located in a loop just above the active site approximately 10 Å from the active site zinc ions. Together, our crystal structures of these NDM variants, including the first known structure of NDM‐12, biochemical analyses, and biophysical analyses provide a basis for understanding the roles of key residues surrounding the active site within NDM variants. A better understanding of clinically relevant NDM variants is essential in the fight against MBL‐mediated antibiotic resistance. Our structural, biochemical, and biophysical studies will help to inform drug design efforts to develop new MBL inhibitors. Support or Funding Information The authors acknowledge financial support from the US National Institutes of Health (Award No. R01 GM111926 to RCP, DLT, MWC, and WF) and institutional support from Miami University through the Robert H. and Nancy J. Blayney Professorship (to RCP). The Advanced Light Source is supported by the US Department of Energy under contract number DE‐AC03‐76SF00098 at Lawrence Berkeley National Laboratory.},

  doi          = {10.1096/fasebj.31.1_supplement.777.21},

  journal      = {FASEB Journal},

  number       = S1,

  volume       = 31,

  place        = {United States},

  year         = {Wed Oct 03 00:00:00 EDT 2018},

  month        = {Wed Oct 03 00:00:00 EDT 2018}

}

Copy to clipboard

Journal Article:

Free Publicly Available Full Text

Accepted Manuscript (Publisher)

Publisher's Version of Record
https://doi.org/10.1096/fasebj.31.1_supplement.777.21

Other availability

Search WorldCat to find libraries that may hold this journal

Save / Share:

Export Metadata

Save to My Library

Similar Records in DOE PAGES and OSTI.GOV collections:

Structural and biochemical analysis of the metallo-β-lactamase L1 from emerging pathogen Stenotrophomonas maltophilia revealed the subtle but distinct di-metal scaffold for catalytic activity

Journal Article Kim, Youngchang ; Maltseva, Natalia ; Wilamowski, Mateusz ; ... - Protein Science

Emergence of Enterobacteriaceae harboring metallo-beta-lactamases (MBL) has raised global threats due to their broad antibiotic resistance profiles and the lack of effective inhibitors against them. We have been studied one of the emerging environmental MBL, the L1 from Stenotrophomonas maltophilia K279a. We determined several crystal structures of L1 complexes with three different classes of beta-lactam antibiotics (penicillin G, moxalactam, meropenem, and imipenem), with the inhibitor captopril and different metal ions (Zn⁺², Cd⁺², and Cu⁺²). All hydrolyzed antibiotics and the inhibitor were found binding to two Zn⁺² ions mainly through the opened lactam ring and some hydrophobic interactions with the bindingmore »« less
Cited by 17
https://doi.org/10.1002/pro.3804
Heteroaryl Phosphonates as Noncovalent Inhibitors of Both Serine- and Metallocarbapenemases

Journal Article Pemberton, Orville A. ; Jaishankar, Priyadarshini ; Akhtar, Afroza ; ... - Journal of Medicinal Chemistry

Gram-negative pathogens expressing serine β-lactamases (SBLs) and metallo-β-lactamases (MBLs), especially those with carbapenemase activity, threaten the clinical utility of almost all β-lactam antibiotics. In this study we describe the discovery of a heteroaryl phosphonate scaffold that exhibits noncovalent cross-class inhibition of representative carbapenemases, specifically the SBL KPC-2 and the MBLs NDM-1 and VIM-2. The most potent lead, compound 16, exhibited low nM to low μM inhibition of KPC-2, NDM-1, and VIM-2. Compound 16 potentiated imipenem efficacy against resistant clinical and laboratory bacterial strains expressing carbapenemases while showing some cytotoxicity toward human HEK293T cells only at concentrations above 100 μg/mL. Complexmore »« less
Cited by 17
https://doi.org/10.1021/acs.jmedchem.9b00728

Full Text Available
Shifting Sands: Modulating Interactions and Outputs of the CHIP‐Hsp70/HOP‐Hsp70 Protein Quality Control Complexes

Journal Article Page, Richard C. ; Zhang, Huaqun ; Cottingim, Kelsey ; ... - FASEB Journal

The 70‐kilodalton heat shock protein (Hsp70) interacts with tetratricopeptide repeat (TPR) domains of CHIP (C‐terminus of Hsp70 Interacting Protein) and HOP (Hsp Organizing Protein) using a two‐carboxylate clamp binding mechanism. Differential affinity for the TPR domains of CHIP and HOP are achieved through interactions with the six residues preceding the carboxy‐terminal Asp ₆₄₁ of Hsp70. In particular, phosphorylation of Hsp70 Thr ₆₃₆ produces a dramatic swing in binding affinities that shift preferential formation of CHIP/Hsp70 complexes to preferential formation of HOP/Hsp70 complexes. Given that CHIP‐Hsp70 complexes promote ubiquitin‐mediated degradation of Hsp70‐chaperoned clients while HOP‐Hsp70 complexes promote refolding of Hsp70‐chaperoned clients,more »« less
https://doi.org/10.1096/fasebj.31.1_supplement.915.6
Clinical Variants of New Delhi Metallo-β-Lactamase Are Evolving To Overcome Zinc Scarcity

Journal Article Stewart, Alesha C. ; Bethel, Christopher R. ; VanPelt, Jamie ; ... - ACS Infectious Diseases

Use and misuse of antibiotics have driven the evolution of serine β-lactamases to better recognize new generations of β-lactam drugs, but the selective pressures driving evolution of metallo-β-lactamases are less clear. Here in this paper, we present evidence that New Delhi metallo-β-lactamase (NDM) is evolving to overcome the selective pressure of zinc(II) scarcity. Studies of NDM-1, NDM-4 (M154L), and NDM-12 (M154L, G222D) demonstrate that the point mutant M154L, contained in 50% of clinical NDM variants, selectively enhances resistance to the penam ampicillin at low zinc(II) concentrations relevant to infection sites. Each of the clinical variants is shown to be progressivelymore »« less
Cited by 38
https://doi.org/10.1021/acsinfecdis.7b00128

Full Text Available
A close look onto structural models and primary ligands of metallo-β-lactamases

Journal Article Raczynska, Joanna E. ; Shabalin, Ivan G. ; Minor, Wladek ; ... - Drug Resistance Updates

β-Lactamases are hydrolytic enzymes capable of opening the β-lactam ring of antibiotics such as penicillin, thus endowing the bacteria that produce them with antibiotic resistance. Of particular medical concern are metallo-β-lactamases (MBLs), with an active site built around coordinated Zn cations. MBLs are pan-reactive enzymes that can break down almost all classes of β-lactams, including such last-resort antibiotics as carbapenems. They are not only broad-spectrum-reactive but are often plasmid-borne (e.g., the New Delhi enzyme, NDM), and can spread horizontally even among unrelated bacteria. Acquired MBLs are encoded by mobile genetic elements, which often include other resistance genes, making the microbiologicalmore »« less
Cited by 39
https://doi.org/10.1016/j.drup.2018.08.001

Full Text Available

Similar Records