Last Updated: 02/09/2025
Structural studies of Plasmodium PIR proteins and their interactions with human inhibitory immune receptors
Objectives
To understand the role of the most commonly found protein on the surface of the malaria-infected blood cells, showing us how they modulate the immune system and revealing whether we can find an invariant site on these molecules which we can target therapeutically to as part of our quest to destroy this deadly parasite.
This project will use the expertise of the Higgins group at structural and functional characterisation of host-parasite interactions. This will be complemented by collaborators who bring expertise in genomic analysis, analysis of immune cell function and animal models of malaria infection.
PIR proteins, human receptors and antibody Fab fragments will expressed in a secreted form from mammalian cell culture systems. Monoclonal antibodies may be generated by immunizing mice, fusing splenocytes, selecting clonal hybridomas in semi-solid media and screening reactivity by ELISA. Fab fragments will be produced by papain cleavage. Proteins, antibodies and protein complexes will be purified by standard chromatography methods. Protein-protein interactions will be assessed by surface plasmon resonance, microscale thermophoresis and isothermal titration calorimetry. Structures will be determined by x-ray crystallography using molecular replacement, sulphur SAD and heavy metal-based approaches for phasing. Binding surfaces will be determined using hydrogen-deuterium exchange mass spectrometry and confirmed using mutagenesis. Sequence analysis will be conducted using MEME analysis with sequence determinants identified from structural features, or from sequences identified as containing ligand binding determinants.
Jan 2020 — Dec 2022
$591,170
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