University of Groningen
Analysis of the ATCase catalysis within the amino acid metabolism of the human malaria
parasite Plasmodium falciparum
Bosch, Soraya Soledad
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Publication date: 2019
Link to publication in University of Groningen/UMCG research database
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Bosch, S. S. (2019). Analysis of the ATCase catalysis within the amino acid metabolism of the human malaria parasite Plasmodium falciparum. University of Groningen.
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CHAPTER 2
JUSTIFICATION AND OBJECTIVES
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Malaria, caused by Plasmodium spp., remains with more than 400.000 deaths per year one of the most severe diseases of our time. The few existing antimalarial drugs are losing their efficacy due to the worldwide spreading of parasite’s drug resistance. Therefore the discovery of new targets to interfere with is of the highest importance. However, an intensive knowledge and understanding of the parasite’s biochemistry is required to interfere with the proliferation of the deadly pathogen.
This work is focusing on the plasmodial aspartate carbamoyltransferase, since the pyrimidines are important metabolites during proliferation. The aim of this thesis is to evaluate the biological role of PfATCase in vitro and at a cellular level, using transgenically modified parasites and eventually, to validate it as a drug target against P. falciparum. Further structural information was obtained by crystallisation trials, which were carried out in collaboration with Dr. Matthew R. Groves and Prof. Alexander Dömling at RUG, The Netherlands. To achieve a deeper knowledge of the pyrimidine pathway of the parasite this work had the following objectives:
(I) Cloning of the open reading frame of aspartate carbamoyltransferase
(II) Biochemical characterisation of the recombinant protein by analysis of SDS-PAGE and Western Blot
(III) Structural characterization via crystallization – Performed at RUG
(IV) Evaluation of the effect of the overexpression and absence of the ATCase activity on the viability of P. falciparum via transgenic parasites
(V) Verification of the respective protein expression via Western-blot analysis and transcript level by qRT-PCR
(VI) Drug assay in vitro and in vivo of synthetic compounds for PfATCase