Altmetric
A high-throughput in vitro translation screen towards discovery of novel antimalarial protein translation inhibitors
Publication available at: | http://biorxiv.org/cgi/content/short/248740v1 |
---|
Title: | A high-throughput in vitro translation screen towards discovery of novel antimalarial protein translation inhibitors |
Authors: | Baumann, H Matthews, H Li, M Hu, JJ Willison, K Baum, J |
Item Type: | Working Paper |
Abstract: | Drugs that target protein synthesis are well-validated for use as antimicrobials, yet specific high throughput (HTP) methods to screen for those targeting malaria are lacking. Here, we have developed a cell free in vitro translation (IVT) assay for the human malaria parasite, Plasmodium falciparum, which reconstitutes the native parasite protein translation machinery. Combining clarified IVT lysate with a click beetle luciferase reporter gene fused to untranslated regions of Pf histidine-rich proteins (hrp)-2 and 3, the HTP IVT assay accurately reports protein translation in a 384-well plate format using a standard spectrofluorometer. We validate the assay as effective in detecting compounds targeting the ribosome, ribosome co-factors (elongation factor 2) and cytosolic tRNA synthetases as well as its ability to find translation inhibitors in a blind screen using a high-density assay format amenable for high throughput. This demonstrates an ability to reconstitute the breadth of the parasite eukaryotic protein translation machinery in vitro and use it as a powerful platform for antimalarial drug discovery. |
Issue Date: | 16-Jan-2018 |
URI: | http://hdl.handle.net/10044/1/68424 |
DOI: | https://dx.doi.org/10.1101/248740 |
Publisher: | BioRxiv |
Copyright Statement: | The copyright holder for this preprint is the author/funder, who has granted bioRxiv a license to display the preprint in perpetuity. It is made available under a CC-BY-NC-ND 4.0 International license. |
Sponsor/Funder: | Wellcome Trust |
Funder's Grant Number: | 105686 |
Open Access location: | http://biorxiv.org/cgi/content/short/248740v1 |
Appears in Collections: | Chemistry Biological and Biophysical Chemistry |