A High-Throughput Inhibitor Screen Targeting CLAG3 Export and Membrane Insertion on Human Erythrocytes Infected with...

Area of Science:

• Malariology
• Parasitology
• Molecular Biology

Background:

• * Plasmodium falciparum, the human malaria parasite, remodels its host erythrocyte for intracellular growth.
• * The parasite exports proteins, including CLAG3, to the erythrocyte membrane, forming the plasmodial surface anion channel (PSAC) for nutrient acquisition.

Purpose of the Study:

• * To explore CLAG3 protein trafficking mechanisms.
• * To develop therapies blocking host cell remodeling by screening for inhibitors of CLAG3 trafficking and membrane insertion.

Main Methods:

• * A split NanoLuc reporter system was employed for high-throughput screening.
• * Approximately 52,000 small molecules were screened.
• * Secondary and tertiary screens were used to filter hits and assess nonspecific toxicity.

Main Results:

• * 65 chemically diverse hits were identified.
• * No bona fide inhibitors of CLAG3 host membrane insertion were found in this screen.
• * The study established a screening framework for future, larger screens.

Conclusions:

• * The developed screening framework can be used for larger screens to identify novel export inhibitors.
• * Such inhibitors could provide insights into Plasmodia host cell remodeling.
• * Novel inhibitors may lead to therapies targeting parasite proteins essential for survival.