scholarly journals Novel inhibitors of Mycobacterium tuberculosis GuaB2 identified by a target based high-throughput phenotypic screen

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Jonathan A. G. Cox ◽  
Grace Mugumbate ◽  
Laura Vela-Glez Del Peral ◽  
Monika Jankute ◽  
Katherine A. Abrahams ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
Phenotypic Screen

scholarly journals A high-throughput whole cell screen to identify inhibitors of Mycobacterium tuberculosis

2018 ◽  
Author(s):  
Juliane Ollinger ◽  
Anuradha Kumar ◽  
David M. Roberts ◽  
Mai A. Bailey ◽  
Allen Casey ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Growth Inhibition ◽  
High Throughput ◽  
Whole Cell ◽  
Fluorescent Reporters ◽  
Recombinant Strains ◽  
Phenotypic Screen ◽  
Novel Drugs

AbstractTuberculosis is a disease of global importance for which novel drugs are urgently required. We developed a whole-cell phenotypic screen which can be used to identify inhibitors of Mycobacterium tuberculosis growth. We used recombinant strains of virulent M. tuberculosis which express far-red fluorescent reporters and used fluorescence to monitor growth in vitro. We optimized our high throughput assays using both 96-well and 384-well plates; both formats gave assays which met stringent reproducibility and robustness tests. We screened a compound set of 1105 chemically diverse compounds previously shown to be active against M. tuberculosis and identified primary hits which showed ≥ 90% growth inhibition. We ranked hits and identified three chemical classes of interest – the phenoxyalkylbenzamidazoles, the benzothiophene 1–1 dioxides, and the piperidinamines. These new compound classes may serve as starting points for the development of new series of inhibitors that prevent the growth of M. tuberculosis. This assay can be used for further screening, or could easily be adapted to other strains of M. tuberculosis.

scholarly journals A high-throughput screening assay based on automated microscopy for monitoring antibiotic susceptibility of Mycobacterium tuberculosis phenotypes

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Sadaf Kalsum ◽  
Blanka Andersson ◽  
Jyotirmoy Das ◽  
Thomas Schön ◽  
Maria Lerm
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Live Cell Imaging ◽  
Cell Imaging ◽  
Imaging System ◽  
Aggregate Size ◽  
Live Cell ◽  
Screening Assay ◽  
High Throughput Screening Assay

Abstract Background Efficient high-throughput drug screening assays are necessary to enable the discovery of new anti-mycobacterial drugs. The purpose of our work was to develop and validate an assay based on live-cell imaging which can monitor the growth of two distinct phenotypes of Mycobacterium tuberculosis and to test their susceptibility to commonly used TB drugs. Results Both planktonic and cording phenotypes were successfully monitored as fluorescent objects using the live-cell imaging system IncuCyte S3, allowing collection of data describing distinct characteristics of aggregate size and growth. The quantification of changes in total area of aggregates was used to define IC50 and MIC values of selected TB drugs which revealed that the cording phenotype grew more rapidly and displayed a higher susceptibility to rifampicin. In checkerboard approach, testing pair-wise combinations of sub-inhibitory concentrations of drugs, rifampicin, linezolid and pretomanid demonstrated superior growth inhibition of cording phenotype. Conclusions Our results emphasize the efficiency of using automated live-cell imaging and its potential in high-throughput whole-cell screening to evaluate existing and search for novel antimycobacterial drugs.

scholarly journals High throughput screening of a library based on kinase inhibitor scaffolds against Mycobacterium tuberculosis H37Rv

Tuberculosis ◽  
2012 ◽  
Vol 92 (1) ◽  
pp. 72-83 ◽  
Author(s):  
Robert C. Reynolds ◽  
Subramaniam Ananthan ◽  
Ellen Faaleolea ◽  
Judith V. Hobrath ◽  
Cecil D. Kwong ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Kinase Inhibitor ◽  
Tuberculosis H37rv ◽  
Mycobacterium Tuberculosis H37rv ◽  
Inhibitor Scaffolds

Selection and optimization of hits from a high-throughput phenotypic screen againstTrypanosoma cruzi

2013 ◽  
Vol 5 (15) ◽  
pp. 1733-1752 ◽  
Author(s):  
Martine Keenan ◽  
Paul W Alexander ◽  
Jason H Chaplin ◽  
Michael J Abbott ◽  
Hugo Diao ◽  
...  
Keyword(s):  
High Throughput ◽  
Phenotypic Screen

scholarly journals High Throughput Phenotypic Selection of Mycobacterium tuberculosis Mutants with Impaired Resistance to Reactive Oxygen Species Identifies Genes Important for Intracellular Growth

PLoS ONE ◽  
2013 ◽  
Vol 8 (1) ◽  
pp. e53486 ◽  
Author(s):  
Olga Mestre ◽  
Raquel Hurtado-Ortiz ◽  
Tiago Dos Vultos ◽  
Amine Namouchi ◽  
Mena Cimino ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
Reactive Oxygen ◽  
Phenotypic Selection ◽  
Oxygen Species ◽  
Intracellular Growth ◽  
Selection Of

scholarly journals Construction of a series of vectors for high throughput cloning and expression screening of membrane proteins from Mycobacterium tuberculosis

2008 ◽  
Vol 8 (1) ◽  
pp. 51 ◽  
Author(s):  
Huajun Qin ◽  
Jian Hu ◽  
Yuanzhi Hua ◽  
Shridhar V Challa ◽  
Timothy A Cross ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Membrane Proteins ◽  
High Throughput ◽  
Cloning And Expression ◽  
Expression Screening

scholarly journals Utility of mycobacterial interspersed repetitive unit typing for differentiating Mycobacterium tuberculosis isolates in Wuhan, China

2007 ◽  
Vol 56 (9) ◽  
pp. 1219-1223 ◽  
Author(s):  
Hui Han ◽  
Fang Wang ◽  
Yong Xiao ◽  
Yi Ren ◽  
Yanjie Chao ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Molecular Epidemiology ◽  
High Throughput ◽  
Discriminatory Power ◽  
Fast Method ◽  
Beijing Family ◽  
Wuhan City ◽  
Preliminary Screening ◽  
Discriminatory Index

Mycobacterial interspersed repetitive unit (MIRU) typing has been found to allow rapid, reliable, high-throughput genotyping of Mycobacterium tuberculosis, and may represent a feasible approach to study M. tuberculosis molecular epidemiology. To evaluate the use of MIRU typing in discriminating M. tuberculosis strains, isolates from 105 patients in Wuhan City, China, were genotyped by this method as compared to spoligotyping. MIRU typing identified 55 types that defined 21 clusters and 34 unique isolates. The discriminatory power was high [Hunter–Gaston discriminatory index (HGDI), 0.97]. Spoligotyping showed that 86 (81.9 %) of 105 isolates belonged to the Beijing family genotype. For Beijing family and non-Beijing strains, the discriminatory power of MIRU was high (HGDI, 0.95 and 0.98, respectively). Among the alleles of the MIRU loci for the Beijing family, only locus 26 was highly discriminative, but for non-Beijing strains, loci 10, 16 and 26 were highly discriminative. MIRU typing is a simple and fast method which may be used for preliminary screening of M. tuberculosis isolates in China.

scholarly journals Development of a Simple Assay Protocol for High-Throughput Screening of Mycobacterium tuberculosis Glutamine Synthetase for the Identification of Novel Inhibitors

2005 ◽  
Vol 10 (7) ◽  
pp. 725-729 ◽  
Author(s):  
Upasana Singh ◽  
Vinita Panchanadikar ◽  
Dhiman Sarkar
Keyword(s):  
Escherichia Coli ◽  
Mycobacterium Tuberculosis ◽  
Glutamine Synthetase ◽  
Small Molecules ◽  
High Throughput ◽  
High Throughput Screening ◽  
Coli Strain ◽  
Compound Library ◽  
Essential Enzyme ◽  
Assay Protocol

Mycobacterium tuberculosis glutamine synthetase (GS) is an essential enzyme involved in the pathogenicity of the organism. The screening of a compound library using a robust high-throughput screening (HTS) assay is currently thought to be the most efficient way of getting lead molecules, which are potent inhibitors for this enzyme. The authors have purified the enzyme to a >90% level from the recombinant Escherichia coli strain YMC21E, and it was used for partial characterization as well as standardization experiments. The results indicated that the Kmof the enzyme for L-glutamine and hydroxylamine were 60 mM and 8.3 mM, respectively. The Km for ADP, arsenate, and Mn2+ were 2 [.proportional]M, 5 [.proportional]M, and 25 [.proportional]M, respectively. When the components were adjusted according to their Km values, the activity remained constant for at least 3 h at both 25° C and 37° C. The Z′ factor determined in microplate format indicated robustness of the assay. When the signal/noise ratios were determined for different assay volumes, it was observed that the 200-[.proportional]l volume was found to be optimum. The DMSO tolerance of the enzyme was checked up to 10%, with minimal inhibition. The IC50 value determined for L-methionine S-sulfoximine on the enzyme activity was 3 mM. Approximately 18,000 small molecules could be screened per day using this protocol by a Beckman Coulter HTS setup.

ChemInform Abstract: Discovery of Novel Inhibitors Targeting the Mycobacterium tuberculosis O-Acetylserine Sulfhydrylase (CysK1) Using Virtual High-Throughput Screening.

ChemInform ◽  
2013 ◽  
Vol 44 (28) ◽  
pp. no-no
Author(s):  
Variam Ullas Jean Kumar ◽  
Oemer Poyraz ◽  
Shalini Saxena ◽  
Robert Schnell ◽  
Perumal Yogeeswari ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening

scholarly journals Mycobacterium tuberculosis virulence inhibitors discovered by Mycobacterium marinum high-throughput screening

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Hasan Tükenmez ◽  
Isabel Edström ◽  
Ramesh Ummanni ◽  
Stina Berglund Fick ◽  
Charlotta Sundin ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Mycobacterium Marinum
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