Plasmodium falciparum exports over 450 proteins to the infected erythrocyte that remodel the cell and mediate virulence. The majority of proteins are exported via recognition and cleavage of the Plasmodium export element (PEXEL) in the N-terminus of exported proteins by the parasite ER-resident protease Plasmepsin V. The gene encoding Plasmepsin V is refractory to deletion, suggesting it is essential for parasite survival. We have generated a small molecule inhibitor that potently blocks the activity of Plasmepsin V isolated from the two most lethal parasites, P. falciparum and P. vivax. Treatment of P. falciparum-infected erythrocytes with the inhibitor caused accumulation of uncleaved PEXEL precursors within the parasite, impairing export to the host cell and killing parasites at the early trophozoite stage. This establishes that Plasmepsin V function and specificity is conserved across Plasmodium and is essential for protein export and erythrocyte remodeling that sustains parasites as they develop in the host. This validates the protease as an important antimalarial drug target. Plasmepsin V inhibitors are now being employed to study a similar PEXEL export mechanism in the related Apicomplexan parasite, Toxoplasma gondii.