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Malaria based proteomics of erythrocyte surface proteins

Malaria based proteomics of erythrocyte surface proteins. Kim Baron University of Pretoria, Department of Pharmacology Supervisor: Dr Duncan Cromarty Co-supervisor: Dr Stoyan Stoychev. Introduction. Few diseases have had same global health & economic impacts of malaria

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Malaria based proteomics of erythrocyte surface proteins

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  1. Malaria based proteomics of erythrocyte surface proteins Kim Baron University of Pretoria, Department of Pharmacology Supervisor: Dr Duncan Cromarty Co-supervisor: Dr Stoyan Stoychev

  2. Introduction • Few diseases have had same global health & economic impacts of malaria • 300 million people affected globally each year & 0.7-2.7 million annual deaths • Caused by protozoan parasite Plasmodium (falciparum) most severe form humans • Discovered 100 years ago  stage specific differential protein expression

  3. Control of malaria uses several approaches • Improve diagnosis • Improve prophylactic chemotherapy • Integrated vector control • Prevalence malaria evident worldwide • Drug-resistant parasites • Insecticide-resistant vectors • New malaria drugs are expensive & limited in supply • Safe, effective & affordable treatment  critical global public health priority

  4. The life cycle of P. falciparum in the human host & the Anopheles mosquito vector

  5. Merozoite invasion of erythrocytes A- Recognition C + D- Tight junction formation/movement B- Reorientation E- Internalisation

  6. Merozoite invasion of RBC’s is highly complex  cascade of specific molecular interactions • Complete receptor-ligand interactions unknown • Invasion is central & essential in development of malaria  speed & efficiency contributed to evolutionary success & parasitemia • Inhibition of parasite invasion before manifestation of clinical symptoms  Advantage

  7. Aim • To identify which erythrocyte surface proteins act as receptors during the initial recognition and binding of the Plasmodium falciparum merozoite to the erythrocyte during the invasion process of the malaria parasite

  8. Method Overview

  9. Results: SDS-PAGE

  10. 2-DE Band 3?? ASB (amidosulfobetaine)-14  zwitterionic detergent

  11. ASB-14 SDS-PAGE Comparison * * Erythrocyte ghosts: absence of ASB-14 Erythrocyte ghosts: sample prep. with ASB-14

  12. Improvements • ASB-14 utilisation with adjusted voltage settings • Active sample loading • 10% T gel instead of 4-20% T

  13. Conclusion • Plasmodium falciparum  complex & “intelligent” • RBC’s are abundant & fragile • Avoids damaging RBC • Protected from immune system • To potentially stop deadly parasite  new innovative therapies • Determining exact R-L during invasion  discovery of new key targets for drug & vaccine based antimalarial strategies

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