Peptide Inhibitors of the <i>Escherichia coli</i> DsbA Oxidative Machinery Essential for Bacterial Virulence

Duprez, Wilko; Premkumar, Lakshmanane; Halili, Maria A.; Lindahl, Fredrik; Reid, Robert C.; Fairlie, David P.; Martin, Jennifer L.

doi:10.1021/jm500955s

Cited by 48 publications

(71 citation statements)

References 53 publications

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“…Our finding that DsbA C33A inhibits VKORc1 in the presence of DsbA wt demonstrates the feasibility of finding a polypeptidederived inhibitor of VKORc1, analogously to recent efforts aimed at designing peptide inhibitors of the DsbB-DsbA complex (32,33). In addition, our agar-based blue/white screen reported previously (14) provides a simple high-throughput screen for potential small-molecule anticoagulants against VKORc1.…”

Section: Discussionsupporting

confidence: 71%

Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase

Hatahet

Blazyk

Martineau

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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Functional overexpression of polytopic membrane proteins, particularly when in a foreign host, is often a challenging task. Factors that negatively affect such processes are poorly understood. Using the mammalian membrane protein vitamin K epoxide reductase (VKORc1) as a reporter, we describe a genetic selection approach allowing the isolation of Escherichia coli mutants capable of functionally expressing this blood-coagulation enzyme. The isolated mutants map to components of membrane protein assembly and quality control proteins YidC and HslV. We show that changes in the VKORc1 sequence and in the YidC hydrophilic groove along with the inactivation of HslV promote VKORc1 activity and dramatically increase its expression level. We hypothesize that such changes correct for mismatches in the membrane topogenic signals between E. coli and eukaryotic cells guiding proper membrane integration. Furthermore, the obtained mutants allow the study of VKORc1 reaction mechanisms, inhibition by warfarin, and the high-throughput screening for potential anticoagulants.membrane protein assembly | YidC | VKORc1 | anticoagulants

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Section: Discussionsupporting

confidence: 71%

Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase

Hatahet

Blazyk

Martineau

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

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“…One of the compounds was shown to reduce bacterial motility, but it did not affect cell growth (88). Using a virtual screening approach, Duprez et al (89) identified a set of noncovalent inhibitors of DsbA that exhibit inhibitory activity at millimolar concentrations (89). While the first two approaches are aimed at DsbA, the Beckwith group (90) employed a cell-based screening method to find compounds that target DsbB and revealed several inhibitors with a pyridazinone core.…”

Section: Adhesive Pilus Proteins Reveal Oxidative Protein-folding Patmentioning

confidence: 99%

Disulfide-Bond-Forming Pathways in Gram-Positive Bacteria

Reardon-Robinson

Ton‐That

2016

J Bacteriol

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Disulfide bonds are important for the stability and function of many secreted proteins. In Gram-negative bacteria, these linkages are catalyzed by thiol-disulfide oxidoreductases (Dsb) in the periplasm. Protein oxidation has been well studied in these organisms, but it has not fully been explored in Gram-positive bacteria, which lack traditional periplasmic compartments. Recent bioinformatics analyses have suggested that the high-GC-content bacteria (i.e., actinobacteria) rely on disulfide-bond-forming pathways. In support of this, Dsb-like proteins have been identified in Mycobacterium tuberculosis, but their functions are not known. Actinomyces oris and Corynebacterium diphtheriae have recently emerged as models to study disulfide bond formation in actinobacteria. In both organisms, disulfide bonds are catalyzed by the membrane-bound oxidoreductase MdbA. Remarkably, unlike known Dsb proteins, MdbA is important for pathogenesis and growth, which makes it a potential target for new antibacterial drugs. This review will discuss disulfide-bond-forming pathways in bacteria, with a special focus on Gram-positive bacteria.

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“…Using the crystal structure of the DsbA-DsbB complex [127], a peptide of seven amino acids corresponding to a loop of DsbB involved in docking with DsbA was identified and found to bind to EcDsbA with low micromolar affinity (K d = 13.1 ± 0.4 μM). Further engineering of this peptide resulted in a new peptide with greater affinity (K d = 5.7 ± 0.4 μM) that also exhibited fairly potent inhibition of EcDsbA oxidase activity (IC 50 = 8.8 ± 1.1 μM) [128]. The studies described herein clearly show that the DsbA-DsbB protein system is an attractive and tractable target for novel antibiotic development.…”

Section: Dsb Enzymes As Novel Antimicrobial Targetsmentioning

confidence: 72%

From Biology to Biotechnology: Disulfide Bond Formation in <i>Escherichia coli</i>

Landgraf¹,

Ren²,

Masuch³

et al. 2017

≪i>Escherichia Coli

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Disulfide bonds formed between a pair of oxidized cysteines are important to the structural integrity and proper folding of many proteins. Accordingly, Nature has evolved several systems for the genesis and maintenance of such bonds. Beginning with the discovery of protein disulfide isomerase, which provided the first evidence for enzyme-catalyzed disulfide-bond formation, many years of research have resulted in the explication of the complex network of electron transport pathways needed for this process. Herein, we take a historical approach in describing the elucidation of disulfide-bond formation in E. coli. We frame this topic in the context of genome sequencing eras. The first section describes the discovery of eukaryotic protein disulfide isomerase and the subsequent research that followed from the early 1960s to the early 1990s, a time period we have named the pre-genomic sequencing era. The second section details the renaissance in research on disulfide-bond formation in the periplasm of prokaryotes, fueled by bacterial genetic screens and the development of genomic sequencing technology. Accordingly, we have named this section the genomic sequencing era, which ranges from the early 1990s to approximately 2010. The final section outlines the use of bacterial genetic screens to select for new oxidoreductase enzymes and their potential uses in biotechnological and pharmaceutical applications. This era we have dubbed the post-genomic sequencing era, and we envision it to represent the future of research on oxidative folding.

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Peptide Inhibitors of the Escherichia coli DsbA Oxidative Machinery Essential for Bacterial Virulence

Cited by 48 publications

References 53 publications

Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase

Altered Escherichia coli membrane protein assembly machinery allows proper membrane assembly of eukaryotic protein vitamin K epoxide reductase

Disulfide-Bond-Forming Pathways in Gram-Positive Bacteria

From Biology to Biotechnology: Disulfide Bond Formation in <i>Escherichia coli</i>

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