Trehalose-6-phosphate synthase from the cat flea Ctenocephalides felis and Drosophila melanogaster: gene identification, cloning, heterologous functional expression and identification of inhibitors by high throughput screening

2012 ◽  
Vol 21 (4) ◽  
pp. 456-471 ◽  
Author(s):  
C. Kern ◽  
C. Wolf ◽  
F. Bender ◽  
M. Berger ◽  
S. Noack ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
High Throughput ◽  
High Throughput Screening ◽  
Functional Expression ◽  
Ctenocephalides Felis ◽  
Gene Identification ◽  
Cat Flea ◽  
Phosphate Synthase

A High-Throughput Screening Assay for Simultaneous Selection of Inhibitors of Mycobacterium tuberculosis 1-Deoxy-D-Xylulose-5-Phosphate Synthase (Dxs) or 1-Deoxy-D-Xylulose 5-Phosphate Reductoisomerase (Dxr)

2011 ◽  
Vol 16 (3) ◽  
pp. 303-312 ◽  
Author(s):  
Vaishali Humnabadkar ◽  
Ramesh K. Jha ◽  
Nuzhat Ghatnekar ◽  
Sunita M. De Sousa
Keyword(s):  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
High Throughput Screening ◽  
Isothermal Calorimetry ◽  
Nicotinamide Adenine Dinucleotide Phosphate ◽  
Enzyme Assays ◽  
High Concentration ◽  
High Throughput Screening Assay ◽  
Individual Enzyme ◽  
Phosphate Synthase

1-deoxy-D-xylulose 5-phosphate reductoisomerase (Dxr) is involved in the synthesis of isoprenoids by the methylerythritol phosphate pathway. Dxr is essential in Mycobacterium tuberculosis ( Mtu), absent in humans and amenable to structure-aided design. To further assess the druggability of the enzyme, the energetics of binding of fosmidomycin to Mtu Dxr was studied by isothermal calorimetry. Binding was enhanced by nicotinamide adenine dinucleotide phosphate hydrogen (NADPH) and driven by enthalpy (ΔH –10.2 kcal/mol, ΔS 1.1 cal mol−1K−1). This suggests the possibility of finding novel inhibitors that bind enthalpically, making Dxr an attractive target. The cost of the Dxr substrate, 1-deoxy-D-xylulose-5-phosphate, for high-throughput screening (HTS) is prohibitive. Hence, an HTS assay that couples Dxr to the upstream enzyme 1-deoxy-D-xylulose-5-phosphate synthase (Dxs), also a valid target, was developed. A high concentration of NADPH was used to bias it to detect Dxr inhibitors that bind like fosmidomycin. The assay Z′ was 0.75. It was equally sensitive to inhibitors of Dxs and Dxr, that is, fosmidomycin and fluropyruvate inhibited it with IC50s similar to that in the individual enzyme assays (79 vs 54 nM for fosmidomycin). To distinguish inhibitors of Dxs from Dxr, individual enzyme assays and a microplate thermofluor binding assay were developed. The assay simultaneously screens two targets and is cost-effective.

scholarly journals Drosophila melanogaster as a function-based high-throughput screening model for antinephrolithiasis agents in kidney stone patients

2018 ◽  
Vol 11 (11) ◽  
pp. dmm035873 ◽  
Author(s):  
Sohrab N. Ali ◽  
Thamara K. Dayarathna ◽  
Aymon N. Ali ◽  
Tijani Osumah ◽  
Mohamed Ahmed ◽  
...  
Keyword(s):  
Drosophila Melanogaster ◽  
High Throughput ◽  
Kidney Stone ◽  
High Throughput Screening ◽  
Screening Model

High-throughput screening program for botanical extracts

Planta Medica ◽  
2012 ◽  
Vol 78 (11) ◽  
Author(s):  
L Hingorani ◽  
NP Seeram ◽  
B Ebersole
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Screening Program ◽  
Botanical Extracts

High throughput screening of microbial biodiversity for the discovery of novel cosmeceutical agents

Planta Medica ◽  
2015 ◽  
Vol 81 (16) ◽  
Author(s):  
K Georgousaki ◽  
N DePedro ◽  
AM Chinchilla ◽  
N Aliagiannis ◽  
F Vicente ◽  
...  
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Microbial Biodiversity

High throughput screening to investigate Brazilian Cerrado biome plant extract bank chemical diversity

Planta Medica ◽  
2016 ◽  
Vol 81 (S 01) ◽  
pp. S1-S381
Author(s):  
LS Espindola ◽  
RG Dusi ◽  
KR Gustafson ◽  
J McMahon ◽  
JA Beutler
Keyword(s):  
High Throughput ◽  
Plant Extract ◽  
High Throughput Screening ◽  
Chemical Diversity ◽  
Brazilian Cerrado ◽  
Cerrado Biome
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