Surface Plasmon Resonance – More Than a Screening Technology: Insights in the Binding Mode of σ
            <sup>70</sup>
            :Core Rnap Inhibitors

Hüsecken, Kristina; Hinsberger, Stefan; Elgaher, Walid A. M.; Haupenthal, Jörg; Hartmann, Rolf W.

doi:10.4155/fmc.14.105

Cited by 4 publications

(6 citation statements)

References 38 publications

(55 reference statements)

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“…The solution was incubated on ice for 2 h and mixed carefully every 30 min. The biotinylated CYP121 was purified by size exclusion chromatography using the storage buffer and subsequently stored at −80 °C at a final concentration of 10 μ m …”

Section: Methodsmentioning

confidence: 99%

Biophysical Screening of a Focused Library for the Discovery of CYP121 Inhibitors as Novel Antimycobacterials

et al. 2017

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The development of novel antimycobacterial agents against Mycobacterium tuberculosis (Mtb) is urgently required due to the appearance of multidrug resistance (MDR) combined with complicated long‐term treatment. CYP121 was shown to be a promising novel target for inhibition of mycobacterial growth. In this study, we describe the rational discovery of new CYP121 inhibitors by a systematic screening based on biophysical and microbiological methods. The best hits originating from only one structural class gave initial information about molecular motifs required for binding and activity. The initial screening procedure was followed by mode‐of‐action studies and further biological characterizations. The results demonstrate superior antimycobacterial efficacy and a decreased toxicity profile of our frontrunner compound relative to the reference compound econazole. Due to its low molecular weight, promising biological profile, and physicochemical properties, this compound is an excellent starting point for further rational optimization.

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Section: Methodsmentioning

confidence: 99%

Biophysical Screening of a Focused Library for the Discovery of CYP121 Inhibitors as Novel Antimycobacterials

et al. 2017

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“…In these studies, we were also able to identify a new heterologous electron‐transfer system composed of Etp1fd (516‐618) as ferrodoxin and Arh1A18G as reductase . Biotinylated CYP125 was bound to the streptavidin‐labeled SPR spectroscopy sensor chip …”

Section: Resultsmentioning

confidence: 99%

“…[38,39] Biotinylated CYP125 was bound to the streptavidin-labeledS PR spectroscopys ensor chip. [40] The reference compound Eco was reported to form at ype II complex with the heme iron of CYP125. [7] For initial validation of our SPR spectroscopy technique, we confirmed Eco binding at aconcentration of 100 mm to immobilizedCYP125( Figure S1 in the Supporting Information).…”

Section: Resultsmentioning

confidence: 99%

“…The solution was incubated on ice for 2 h and was mixed carefully every 30 min. Biotinylated CYP121 was purified by a size‐exclusion column by using storage buffer and stored at −80 °C at a final concentration of 10 μ m …”

Section: Methodsmentioning

confidence: 99%

“…Biotinylated CYP121 was purified by as ize-exclusion column by using storage buffer and stored at À80 8Catafinal concentration of 10 mm. [40] Spectroscopic characterization of enzyme activity:F erredoxin Etp1fd (516-618) and ferredoxin reductase Arh1A18G from the fission yeast Schizosaccharomyces pombe were expressed and purified as previously described. [38,39] Functionality of CYP125 and electron transfer was assayed by the occurrence of the characteristic peak at approximately l = 450 nm, related to the reduced, CO-bound heme complex.…”

Section: Biological Methodsmentioning

confidence: 99%

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Discovery and Biophysical Evaluation of First Low Nanomolar Hits Targeting CYP125 of M. tuberculosis

et al. 2016

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Tuberculosis, which is predominantly caused by Mycobacterium tuberculosis (Mtb), is still the most lethal bacterial infection with 1.5 million casualties in 2014. Moreover, the fact that the appearance of resistant mutants and long-term treatment are coupled with economic problems in developing countries hampers an efficient therapy. Interference with the essential cholesterol metabolism of Mtb could be a promising novel strategy to fight Mtb infections. CYP125, a P450 enzyme in Mtb, has been shown to play an important role in this metabolic pathway. For this reason, we used a combined screening approach involving surface plasmon resonance spectroscopy and a heme coordination assay to identify new CYP125 binders by employing a focused P450-inhibitor library. We identified the first hits with high affinity and favorable ligand efficiencies. Furthermore, frontrunner compounds also showed selectivity toward CYP121 specific to Mtb and required for its survival. To date, these are the first compounds targeting CYP125 with low nanomolar affinity.

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Nanopore-Based Sensors for Ligand–Receptor Lead Optimization

Luan

Huynh

Zhou

2015

J. Phys. Chem. Lett.

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Developing a low-cost and high-efficacy method to optimize prescreened or designed drug candidates will facilitate drug discovery and biomedical research in general. Current methods of drug screening usually involve tedious sample preparation and costly biological/chemical assays. Here, through all-atom molecular dynamics simulations, we propose a new drug optimization method, based on the nanopore force spectroscopy, to electrically detect the binding strength between a drug molecule and a target protein. Simulation results demonstrate that the drug-protein complex can be electrophoretically driven into a nanopore, which is followed by the rupture of the complex at a critical biasing voltage. The latter determines the binding strength of the tested drug molecule. It is expected that the application of this single-molecule technology could help to accelerate the drug discovery, particularly for processes of the narrow screening and further lead optimization.

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Surface Plasmon Resonance – More Than a Screening Technology: Insights in the Binding Mode of σ ⁷⁰ :Core Rnap Inhibitors

Abstract: Surface plasmon resonance is a valuable technology in drug design, that has been used in this study to identify and evaluate σ(70):core RNA polymerase inhibitors.

Cited by 4 publications

References 38 publications

Biophysical Screening of a Focused Library for the Discovery of CYP121 Inhibitors as Novel Antimycobacterials

Biophysical Screening of a Focused Library for the Discovery of CYP121 Inhibitors as Novel Antimycobacterials

Discovery and Biophysical Evaluation of First Low Nanomolar Hits Targeting CYP125 of M. tuberculosis

Nanopore-Based Sensors for Ligand–Receptor Lead Optimization

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Surface Plasmon Resonance – More Than a Screening Technology: Insights in the Binding Mode of σ 70 :Core Rnap Inhibitors

Abstract: Surface plasmon resonance is a valuable technology in drug design, that has been used in this study to identify and evaluate σ(70):core RNA polymerase inhibitors.

Cited by 4 publications

References 38 publications

Biophysical Screening of a Focused Library for the Discovery of CYP121 Inhibitors as Novel Antimycobacterials

Biophysical Screening of a Focused Library for the Discovery of CYP121 Inhibitors as Novel Antimycobacterials

Discovery and Biophysical Evaluation of First Low Nanomolar Hits Targeting CYP125 of M. tuberculosis

Nanopore-Based Sensors for Ligand–Receptor Lead Optimization

Contact Info

Product

Resources

About

Surface Plasmon Resonance – More Than a Screening Technology: Insights in the Binding Mode of σ ⁷⁰ :Core Rnap Inhibitors