A Screen to Identify Small Molecule Inhibitors of Protein–Protein Interactions in Mycobacteria

Mai, Deborah; Jones, Jennifer; Rodgers, John W.; Hartman, John L.; Kutsch, Olaf; Steyn, Adrie J. C.

doi:10.1089/adt.2010.0326

Cited by 12 publications

(16 citation statements)

References 84 publications

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“…Indeed, the bacterial or host-pathogen protein interactomes provide invaluable insights for the identification of novel antimicrobial drug targets, as successfully accomplished in cases of many human pathogens 8–10 . A wealth of data involving host- B .…”

Section: Introductionmentioning

confidence: 99%

A protein-protein interaction dictates Borrelial infectivity

Thakur

Sharma

Chao

et al. 2017

Sci Rep

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Two Borrelia burgdorferi interacting proteins, BB0238 and BB0323, play distinct roles in pathogen biology and infectivity although a significance of their interaction remained enigmatic. Here we identified the polypeptide segment essential for BB0238-BB0323 interaction and examined how it supports spirochete infectivity. We show that the interaction region in BB0323 requires amino acid residues 22–200, suggesting that the binding encompasses discontinuous protein segments. In contrast, the interaction region in BB0238 spans only 11 amino acids, residues 120–130. A deletion of these 11 amino acids neither alters the overall secondary structure of the protein, nor affects its stability or oligomerization property, however, it reduces the post-translational stability of the binding partner, BB0323. Mutant B. burgdorferi isolates producing BB0238 lacking the 11-amino acid interaction region were able to persist in ticks but failed to transmit to mice or to establish infection. These results suggest that BB0238-BB0323 interaction is critical for post-translational stability of BB0323, and that this interaction is important for mammalian infectivity and transmission of B. burgdorferi. We show that saturation or inhibition of BB0238-BB0323 interaction could be studied in a luciferase assay, which could be amenable for future identification of small molecule inhibitors to combat B. burgdorferi infection.

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Section: Introductionmentioning

confidence: 99%

A protein-protein interaction dictates Borrelial infectivity

Thakur

Sharma

Chao

et al. 2017

Sci Rep

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“…The wide TRIM susceptibility window between the strains encoding dimerization-proficient and dimerizationimpaired EspR proteins together with the specific correlations between EspR homodimerization, TRIM concentration, and bacterial survival allow for a sensitive and quantitative assay that can be performed in a high-throughput manner (20,26). Screening and characterizing potential antivirulence compounds targeting EspR will be the focus of further research.…”

Section: Discussionmentioning

confidence: 99%

Functional Dissection of Intersubunit Interactions in the EspR Virulence Regulator of Mycobacterium tuberculosis

et al. 2014

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The nucleoid-associated protein EspR, a chromosome organizer, has pleiotropic effects on expression of genes associated with cell wall function and pathogenesis in Mycobacterium tuberculosis. In particular, EspR binds to several sites upstream of the espACD locus to promote its expression, thereby ensuring full function of the ESX-1 secretion system, a major virulence determinant. The N terminus of EspR contains the helix-turn-helix DNA-binding domain, whereas the C-terminal dimerization domain harbors residues involved in intersubunit interactions. While direct binding to DNA appears to be mediated by an EspR dimer-of-dimers, where two helix-turn-helix motifs remain free for long-range interactions, the mechanism of EspR higher-order organization and its impact on chromosome structure and gene expression are not understood. To investigate these processes, we identified seven amino acid residues using molecular dynamics and replaced them with Ala in order to probe interactions at either the dimer or the dimer-of-dimers interfaces. Arg70, Lys72, and Arg101 were important for protein stability and optimal DNA-binding activity. Moreover, the Arg70 mutant showed decreased dimerization in a mycobacterial two-hybrid system. To correlate these defects with higher-order organization and transcriptional activity, we used atomic force microscopy to observe different EspR mutant proteins in complex with the espACD promoter region. In addition, complementation of an M. tuberculosis espR knockout mutant was performed to measure their impact on EspA expression. Our results pinpoint key residues required for EspR function at the dimer (Arg70) and the dimer-of-dimers (Lys72) interface and demonstrate that EspR dimerization and higher-order oligomerization modulate espACD transcriptional activity and hence pathogenesis.

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“…DosRST TCS comprises the DosR response regulator and the DosS and DosT histidine kinases that play an essential role in triggering and maintaining dormancy and 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 Several sets of small molecules have been assayed for their inhibition of DosR activity (63) (64). Several small molecules have been described as DosR inhibitors that bind DosR and inhibit its binding to its target DNA sequence (63).…”

Section: Targeting M Tuberculosis Persistence Through Inhibition Of mentioning

confidence: 99%

Bacterial Histidine Kinases as Novel Antibacterial Drug Targets

et al. 2014

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Bacterial histidine kinases (HKs) are promising targets for novel antibacterials. Bacterial HKs are part of bacterial two-component systems (TCSs), the main signal transduction pathways in bacteria, regulating various processes including virulence, secretion systems and antibiotic resistance. In this review, we discuss the biological importance of TCSs and bacterial HKs for the discovery of novel antibacterials, as well as published TCS and HK inhibitors that can be used as a starting point for structure-based approaches to develop novel antibacterials.

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A Screen to Identify Small Molecule Inhibitors of Protein–Protein Interactions in Mycobacteria

Cited by 12 publications

References 84 publications

A protein-protein interaction dictates Borrelial infectivity

A protein-protein interaction dictates Borrelial infectivity

Functional Dissection of Intersubunit Interactions in the EspR Virulence Regulator of Mycobacterium tuberculosis

Bacterial Histidine Kinases as Novel Antibacterial Drug Targets

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