The Mechanism of Action of Inhibitors of Bacterial Two-component Signal Transduction Systems

Stephenson, Keith; Yamaguchi, Yasuchika; Hoch, James A.

doi:10.1074/jbc.m006633200

Cited by 107 publications

(114 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…However, when tested for their antimicrobial potential in vivo, most of these compounds were not selective for signal transduction pathways, but exerted their effects through multiple mechanisms of action (35), making them unsuitable for any clinical application. Most of these inhibitors were developed by random screening, and most of them were shown to bind to the kinase domain, which might explain their restricted selectivity (36). In this study, we demonstrate that an HPK can be inhibited by structural analogues that behave as antagonists.…”

Section: Ortho-substitutions Of Toluene: Converting An Agonist Molecumentioning

confidence: 86%

Bacterial sensor kinase TodS interacts with agonistic and antagonistic signals

Büsch

Lacal

Martos

et al. 2007

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

105

View full text Add to dashboard Cite

The TodS/TodT two-component system controls expression of the toluene dioxygenase (TOD) pathway for the metabolism of toluene in Pseudomonas putida DOT-T1E. TodS is a sensor kinase that ultimately controls tod gene expression through its cognate response regulator, TodT. We used isothermal titration calorimetry to study the binding of different compounds to TodS and related these findings to their capacity to induce gene expression in vivo. Agonistic compounds bound to TodS and induced gene expression in vivo. Toluene was a powerful agonist, but ortho-substitutions of toluene reduced or abolished in vivo responses, although TodS recognized o-xylene with high affinity. These compounds were called antagonists. We show that agonists and antagonists compete for binding to TodS both in vitro and in vivo. The failure of antagonists to induce gene expression in vivo correlated with their inability to stimulate TodS autophosphorylation in vitro. We propose intramolecular TodS signal transmission, not molecular recognition of compounds by TodS, to be the phenomenon that determines whether a given compound will lead to activation of expression of the tod genes. Molecular modeling identified residues F46, I74, F79, and I114 as being potentially involved in the binding of effector molecules. Alanine substitution mutants of these residues reduced affinities (2-to 345-fold) for both agonistic and antagonistic compounds. Our data indicate that determining the inhibitory activity of antagonists is a potentially fruitful alternative to design specific two-component system inhibitors for the development of new drugs to inhibit processes regulated by two-component systems.histidine kinases ͉ isothermal titration calorimetry ͉ Pseudomonas ͉ two-component systems ͉ aromatic hydrocarbons

show abstract

Section: Ortho-substitutions Of Toluene: Converting An Agonist Molecumentioning

confidence: 86%

Bacterial sensor kinase TodS interacts with agonistic and antagonistic signals

Büsch

Lacal

Martos

et al. 2007

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

105

View full text Add to dashboard Cite

show abstract

“…6,28,29 However, the majority of these inhibitors suffer from excessive protein binding and/or limited bioavailability and have multiple mechanisms of action including bacterial membrane-damaging and surfactant properties 25 and structural alterations of the histidine kinases leading to aggregation. 30 Therefore, these compounds were not as selective as was originally thought and did not exhibit a strong correlation between their IC 50 s against 2-component systems and minimum inhibitory concentration against bacteria. 6,28 Nonspecific interaction of the compounds with proteins unrelated to their molecular target has been suggested as a possible explanation 31 in addition to aggregate formation leading to nonspecific inhibition.…”

Section: Discussionmentioning

confidence: 99%

High-Throughput Microplate Phosphorylation Assays Based on DevR-DevS/Rv2027c 2-Component Signal Transduction Pathway to Screen for Novel Antitubercular Compounds

Saini

Tyagi

2005

SLAS Discovery

View full text Add to dashboard Cite

DevR-DevS (Rv3133c-Rv3132c) and DevR-Rv2027c have been established through their autophosphorylation and phosphotransfer properties to constitute bonafide regulatory 2-component systems of Mycobacterium tuberculosis. DevR has also been shown by others to play a key regulatory role in the expression of M. tuberculosis genes comprising the dormancy regulon. The authors describe high-throughput phosphorylation assays in a microplate format using DevS and Rv2027c histidine kinases and DevR response regulator proteins from M. tuberculosis. The assays were designed to measure [γ-32 P]ATP-dependent autophosphorylation of DevS/Rv2027c and also the phosphotransfer reaction to DevR. First, the optimal reaction conditions were established using the conventional method of radiolabeling the 2-component proteins by [γ-32 P]ATP and followed by gel electrophoresis-based analysis. Next, the assays were converted to a high-throughput format in which the radiolabeled protein retained on a filter using mixed cellulose ester-based 96-well filter plates was analyzed for radioactivity retention by scintillation counting. The utility of these assays to screen for inhibitors is illustrated using 2-mercaptobenzimidazole, ethidium bromide, and EDTA. The high quality and flexibility of these assays will enable their use in high-throughput screening for new antitubercular compounds directed against 2-component systems that comprise a novel target in dormant mycobacteria. (Journal of Biomolecular Screening 2005:215-224)

show abstract

“…Furthermore, DNase I can degrade the extracellular DNA present in the biofilm matrix (Okshevsky et al 2015;Qin et al 2007), interfering with its formation and stability as shown for Bordetella pertussis (Conover et al 2011), Listeria monocytogenes (Harmsen et al, 2010), and Campylobacter jejuni (Brown et al 2015). A bacterial glycoside hydrolase, named Dispersin B, has been isolated from Actinobacillus actinomycetemcomitans, which was shown to disrupt mature Actinobacillus biofilms (Rasko et al 2008;Stephenson et al 2000;Wilke et al 2015;Worthington et al 2013). One prominent example is the small molecular inhibitor Savirin, which reduces the expression of AgrCA-regulated genes in S. aureus, affecting its virulence, but not its survival and has no impact on the commensal Staphylococcus epidermidis (Sully et al 2014).…”

Section: Prevention and Resolution Of Biofilmsmentioning

confidence: 99%

Anti-virulence Strategies to Target Bacterial Infections

Mühlen

Dersch

2015

Current Topics in Microbiology and Immunology

143

139

View full text Add to dashboard Cite

Resistance of important bacterial pathogens to common antimicrobial therapies and the emergence of multidrug-resistant bacteria is increasing at an alarming rate and constitutes one of our greatest challenges in the combat of bacterial infection and accompanied diseases. Given the current shortage of effective drugs, lack of successful prevention measures and only a few new antibiotics in the clinical pipeline demands the development of novel treatment options and alternative antimicrobial therapies. Our increasing understanding of bacterial virulence strategies and the induced molecular pathways of the infectious disease provide novel opportunities to target and interfere with crucial pathogenicity factors or virulence-associated traits of the bacteria while bypassing the evolutionary pressure on the bacterium to develop resistance. In the past decade, numerous new bacterial targets for anti-virulence therapies have been identified, and structure-based tailoring of intervention strategies as well as screening assays for small molecule inhibitors of such pathways were successfully established. In this chapter, we will take a closer look at the bacterial virulence-related factors and processes that present promising targets for antivirulence therapies, recently discovered inhibitory substances and their promises and discuss the challenges and problems that have to be faced.

show abstract

The Mechanism of Action of Inhibitors of Bacterial Two-component Signal Transduction Systems

Cited by 107 publications

References 23 publications

Bacterial sensor kinase TodS interacts with agonistic and antagonistic signals

Bacterial sensor kinase TodS interacts with agonistic and antagonistic signals

High-Throughput Microplate Phosphorylation Assays Based on DevR-DevS/Rv2027c 2-Component Signal Transduction Pathway to Screen for Novel Antitubercular Compounds

Anti-virulence Strategies to Target Bacterial Infections

Contact Info

Product

Resources

About