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Biochemical Characterization of the Apt1/Cdc50 Complex

Project description

Towards novel anti-fungal drugs

Cryptococcus neoformans is a fungal pathogen that can be life-threatening for people with weakened immune systems such as those infected with HIV. The scope of the EU-funded BiCoACC project is to discover novel anti-fungal drugs against Cryptococcus neoformans. Scientists will focus on Apt1 and Cdc50, two fungal proteins critical for fungal virulence known for their role in maintaining phospholipid asymmetry and membrane homeostasis. The biochemical characterisation of these proteins will help identify inhibitors of the Apt1/Cdc50 complex from a chemical library and assess their impact on Cryptococcus neoformans survival and virulence, paving the way for novel anti-fungal agents.

Objective

Cryptococcus neoformans is an opportunist fungal pathogen lethal for patient with AIDS. Treatment options are limited and there is an urgent need for the discovery of anti-fungal drugs. The membrane proteins Apt1 and Cdc50 from C. neoformans were identified as potential drug targets since it is involved in antifungal resistance and pathogen’s virulence. Apt1 belongs to the P4-type ATPase family and together with Cdc50 proteins they are involved in the maintenance of phospholipid asymmetry in biological membranes. In this project I plan to biochemically characterize Apt1/Cdc50 from C. neoformans, to identify inhibitors and functionally impaired mutants of this complex. These in vitro findings will be translated in vivo in C. neoformans cultures. I plan to join the Laboratoire des Protéines et des Systèmes Membranaires (LPSM) at the CEA Paris-Saclay, a world-leading lab on P4-type ATPases, where I will set up protocols for the expression and purification of the Apt1/Cdc50 complex expressed in Saccharomyces cerevisiae. Furthermore, thanks to my expertise in membrane protein reconstitution in liposomes and functional analysis, I will set up an assay to monitor in vitro phospholipid transport, an expertise still lacking at the LPSM. Using purified functional Apt1/Cdc50 complex we will screen a chemical library to identify inhibitors of this complex. In parallel a structure-function study of the complex using directed mutagenesis will be conducted. Finally, in vitro findings will be translated in vivo thanks to a collaboration with Pr Kronstad (University of British Columbia), a world expert on C. neoformans pathophysiology, where C. neoformans cultures are available. This multidisciplinary project will benefit from the expertise of the host institution on P4-ATPases, my knowledge on membrane protein reconstitution in liposomes, and the experience of Pr Kronstad on in vivo study of C. neoformans. This would allow a 3-ways transfer of expertise.

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MSCA-IF - Marie Skłodowska-Curie Individual Fellowships (IF)

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Call for proposal

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(opens in new window) H2020-MSCA-IF-2019

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Coordinator

COMMISSARIAT A L ENERGIE ATOMIQUE ET AUX ENERGIES ALTERNATIVES
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 196 707,84
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RUE LEBLANC 25
75015 Paris
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Ile-de-France Ile-de-France Paris
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Research Organisations
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Total cost

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€ 196 707,84
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