biostudies-literatureWang QQMOST | National Natural Science Foundation of China (NSFC)MOST | National Natural Science Foundation of Chinae0130923https://www.ebi.ac.uk/biostudies/studies/S-EPMC10470541biostudies-literatureUnknown14(4)In the apicomplexans, endocytosed cargos (e.g., hemoglobin) are trafficked to a specialized organelle for digestion. This follows a unique endocytotic process at the micropore/cytostome in these parasites. However, the mechanism underlying endocytic trafficking remains elusive, due to the repurposing of classical endocytic proteins for the biogenesis of apical organelles. To resolve this issue, we have exploited the genetic tractability of the model apicomplexan <i>Toxoplasma gondii</i>, which ingests host cytosolic materials (e.g., green fluorescent protein[GFP]). We determined an association between protein prenylation and endocytic trafficking, and using an alkyne-labeled click chemistry approach, the prenylated proteome was characterized. Genome editing, using clustered regularly intermBioFunctional screening reveals <i>Toxoplasma</i> prenylated proteins required for endocytic trafficking and rhoptry protein sorting.PMC104705413217288732172880Lun ZRWu XTDing HYLiu JYang JLai DHMaity SHe KLong SSun MZhu XQLi YBTang XYHide GTang TDistefano MWang QQfalseFunctional screening reveals <i>Toxoplasma</i> prenylated proteins required for endocytic trafficking and rhoptry protein sorting.In the apicomplexans, endocytosed cargos (e.g., hemoglobin) are trafficked to a specialized organelle for digestion. This follows a unique endocytotic process at the micropore/cytostome in these parasites. However, the mechanism underlying endocytic trafficking remains elusive, due to the repurposing of classical endocytic proteins for the biogenesis of apical organelles. To resolve this issue, we have exploited the genetic tractability of the model apicomplexan <i>Toxoplasma gondii</i>, which ingests host cytosolic materials (e.g., green fluorescent protein[GFP]). We determined an association between protein prenylation and endocytic trafficking, and using an alkyne-labeled click chemistry approach, the prenylated proteome was characterized. Genome editing, using clustered regularly inter2023-01-01T00:00:00Z2023 Aug2025-04-19T20:05:46.174Z2025-04-19T20:05:46.174ZS-EPMC104705413754845210.1128/mbio.01309-23