Microscale Adaptation of <i>In Vitro</i> Transcription/Translation for High‐Throughput Screening of Natural Product Extract Libraries

Lowell, Andrew N.; Santoro, Nicholas; Swaney, Steven; McQuade, Thomas J.; Schultz, Pamela J.; Larsen, Martha J.; Sherman, David H.

doi:10.1111/cbdd.12614

Cited by 11 publications

(11 citation statements)

References 16 publications

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“…Transcription/translation assay was performed as described previously 23 , 24 . Various concentrations of ovatodiolide, kanamycin, erythromycin, and negative controls (0.4% DMSO) were mixed with diluted E .…”

Section: Methodsmentioning

confidence: 99%

Antibacterial activity of ovatodiolide isolated from Anisomeles indica against Helicobacter pylori

Lien

Hung

et al. 2019

Sci Rep

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Helicobacter pylori infection is associated with high incidence of gastric diseases. The extensive therapy of H. pylori infection with antibiotics has increased its resistance rates worldwide. Ovatodiolide, a pure constituent isolated from Anisomeles indica, has been demonstrated to possess bactericidal activity against H. pylori. In this study, ovatodiolide inhibited the growth of both H. pylori reference strain and clinical multidrug-resistant isolates. Docking analysis revealed that ovatodiolide fits into the hydrophobic pocket of a ribosomal protein, RpsB. Furthermore, ovatodiolide inhibited bacterial growth by reducing levels of RpsB, which plays a crucial role in protein translation. Our results demonstrate that ovatodiolide binds to a ribosomal protein and interferes with protein synthesis. This study provides evidence that ovatodiolide has the potential to be developed into a potent therapeutic agent for treating H. pylori infection.

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Section: Methodsmentioning

confidence: 99%

Antibacterial activity of ovatodiolide isolated from Anisomeles indica against Helicobacter pylori

Lien

Hung

et al. 2019

Sci Rep

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“…The significant reduction of the reaction volume to 2 μL allowed the authors to use this in vitro inhibition assay for high-throughput screening [ 21 ]. By means of this assay, 30,000 compounds were tested on 1536-well microplates.…”

Section: In Vitro Methodsmentioning

confidence: 99%

Techniques for Screening Translation Inhibitors

et al. 2016

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The machinery of translation is one of the most common targets of antibiotics. The development and screening of new antibiotics usually proceeds by testing antimicrobial activity followed by laborious studies of the mechanism of action. High-throughput methods for new antibiotic screening based on antimicrobial activity have become routine; however, identification of molecular targets is usually a challenge. Therefore, it is highly beneficial to combine primary screening with the identification of the mechanism of action. In this review, we describe a collection of methods for screening translation inhibitors, with a special emphasis on methods which can be performed in a high-throughput manner.

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“…In 2015, Lowell et al 131 developed a cell‐free miniature bacterial‐coupled transcription/translation assay system. First, the luciferase‐encoding gene was inserted into a plasmid containing the T7 RNA polymerase promoter.…”

Section: Challenges and Advances In Ribosomal Antibiotics And Bacterial Ribosomesmentioning

confidence: 99%

“…Finally, the GraFit program was used to calculate the inhibition rate of each compound. Through this system, over 1300 natural products with over 50% inhibition rate against ribosomal translation at the concentration of 375 μg/ml have been identified for further investigation 131 . Also, by measuring the in vitro translational inhibition ability of a series of lincosamine derivatives, the structure–activity relationship was revealed 132 .…”

Section: Challenges and Advances In Ribosomal Antibiotics And Bacterial Ribosomesmentioning

confidence: 99%

Ribosome‐targeting antibacterial agents: Advances, challenges, and opportunities

Zhang

Bai

et al. 2021

Medicinal Research Reviews

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Ribosomes, which synthesize proteins, are critical organelles for the survival and growth of bacteria. About 60% of approved antibiotics discovered so far combat pathogenic bacteria by targeting ribosomes. However, several issues, such as drug resistance and toxicity, have impeded the clinical use of ribosome‐targeting antibiotics. Moreover, the complexity of the bacteria ribosome structure has retarded the discovery of new ribosome‐targeting agents that are considered as the key to the drug‐resistance and toxicity. To deal with these challenges, efforts such as medicinal chemistry optimization, combination treatment, and new drug delivery system have been developed. But not enough, the development of structural biology and new screening methods bring powerful tools, such as cryo‐electron microscopy technology, advanced computer‐aided drug design, and cell‐free in vitro transcription/translation systems, for the discovery of novel ribosome‐targeting antibiotics. Thus, in this paper, we overview the research on different aspects of bacterial ribosomes, especially focus on discussing the challenges in the discovery of ribosome‐targeting antibacterial drugs and advances made to address issues such as drug‐resistance and selectivity, which, we believe, provide perspectives for the discovery of novel antibiotics.

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Microscale Adaptation of In Vitro Transcription/Translation for High‐Throughput Screening of Natural Product Extract Libraries

Cited by 11 publications

References 16 publications

Antibacterial activity of ovatodiolide isolated from Anisomeles indica against Helicobacter pylori

Antibacterial activity of ovatodiolide isolated from Anisomeles indica against Helicobacter pylori

Techniques for Screening Translation Inhibitors

Ribosome‐targeting antibacterial agents: Advances, challenges, and opportunities

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