Search

Members of the Apicomplexa phylum, including Plasmodium and Toxoplasma, have two types of secretory organelles (micronemes and rhoptries) whose sequential release is essential for invasion and the intracellular lifestyle of these eukaryotes. During invasion, rhoptries inject an array of invasion and virulence factors into the cytoplasm of the host cell, but the molecular mechanism mediating rhoptry exocytosis is unknown. Here we identify a set of parasite specific proteins, termed rhoptry apical surface proteins (RASP) that cap the extremity of the rhoptry. Depletion of RASP2 results in loss of rhoptry secretion and completely blocks parasite invasion and therefore parasite proliferation in both Toxoplasma and Plasmodium. Recombinant RASP2 binds charged lipids and likely contributes to assembling the machinery that docks/primes the rhoptry to the plasma membrane prior to fusion. This study provides important mechanistic insight into a parasite specific exocytic pathway, essential for the establishment of infection. Plasmodium and Toxoplasma parasites rely on rhoptry exocytosis for invasion, but the underlying mechanism is not known. Here, Suarez et al. characterize rhoptry apical surface proteins (RASP) that localize to the rhoptry cap and bind charged lipids, and are essential for rhoptry secretion and invasion. [ABSTRACT FROM AUTHOR]

Parasites of the phylum Apicomplexa are highly successful pathogens of humans and animals worldwide. As obligate intracellular parasites, they have significant energy requirements for invasion and gliding motility that are supplied by various metabolic pathways. Aldolases have emerged as key enzymes involved in these pathways, and all apicomplexans express one or both of fructose 1,6-bisphosphate (F16BP) aldolase and 2-deoxyribose 5-phosphate (dR5P) aldolase (DERA). Intriguingly, Toxoplasma gondii , a highly successful apicomplexan parasite, expresses F16BP aldolase ( Tg ALD1), d5RP aldolase ( Tg DERA), and a divergent dR5P aldolase-like protein ( Tg DPA) exclusively in the latent bradyzoite stage. While the importance of Tg ALD1 in glycolysis is well established and Tg DERA is also likely to be involved in parasite metabolism, the detailed function of Tg DPA remains elusive. To gain mechanistic insight into the function of different T. gondii aldolases, we first determined the crystal structures of Tg ALD1 and Tg DPA. Structural analysis revealed that both aldolases adopt a TIM barrel fold accessorized with divergent secondary structure elements. Structural comparison of Tg ALD1 and Tg DPA with members of their respective enzyme families revealed that, while the active-site residues are conserved in Tg ALD1, key catalytic residues are absent in Tg DPA. Consistent with this observation, biochemical assays showed that, while Tg ALD1 was active on F16BP, Tg DPA was inactive on dR5P. Intriguingly, both aldolases are competent to bind polymerized actin in vitro . Altogether, structural and biochemical analyses of T. gondii aldolase and aldolase-like proteins reveal diverse functionalization of the classic TIM barrel aldolase fold. [ABSTRACT FROM AUTHOR]