Abstract
With 800 thousand deaths each year resulting from more than 225 million cases of malaria worldwide, the increase of drug-resistant parasites and insecticide-resistant Anopheles gambiae mosquitoes has given rise to the call for new approaches in combating malaria. One proposed strategy focuses on the targeting of Plasmodium helicases due to their necessity in all cellular processes, and therefore the survival of the parasite and the infection of their host.This study describes the novel purification and storage strategies of two Plasmodium falciparum helicases; PFEIF4A (formally PFH45) and PFDH60. This resulted in the yield of relatively large amounts of protein that can be used in high-throughput screening of inhibitors and structural-guided drug design in the future.
Moreover, further biochemical and biophysical characterisation has been performed to investigate the specific substrate requirements of each helicase, and the optimal conditions for helicase activity. As well as elucidating information that could provide pathways for specific targeting by both established and novel therapeutic molecules, particularly in the case of PFEIF4A, exposure to temperature just 3 °C above its 37 °C optimum has been shown to have an irreversible and detrimental effect to the proteins activity in vitro.
Preliminary experiments into the proteins’ structures have been performed, as well as experiments exploring the effects of a novel DNA-interactive PBD conjugate and the generation and characterisation of aptamers to PFEIF4A. These experiments have contributed to future investigations in to novel anti-malarial therapeutics and diagnostic platforms.
| Date of Award | Sept 2012 |
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| Original language | English |
| Awarding Institution |
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| Supervisor | Darren Gowers (Supervisor), Andrew Pickford (Supervisor), Anastasia Callaghan (Supervisor), Andrew Pickford (Supervisor), Darren Gowers (Supervisor) & Anastasia Callaghan (Supervisor) |