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Significantly, in mammalian cells CME is responsible for the vast bulk of endocytic flux and has likely undergone multiple adaptations to accommodate specific requirements by individual species. In African trypanosomes, we previously demonstrated that CME is independent of the AP-2 adaptor protein complex, that orthologues to many of the animal and fungal CME protein cohort are absent, and that a novel, trypanosome-restricted protein cohort interacts with clathrin and drives CME. Here, we used a novel cryomilling affinity isolation strategy to preserve transient low-affinity interactions, giving the most comprehensive trypanosome clathrin interactome to date. We identified the trypanosome AP-1 complex, Trypanosoma brucei (Tb)EpsinR, several endosomal SNAREs plus orthologues of SMAP and the AP-2 associated kinase AAK1 as interacting with clathrin. Novel lineage-specific proteins were identified, which we designate TbCAP80 and TbCAP141. Their depletion produced extensive defects in endocytosis and endomembrane system organisation, revealing a novel molecular pathway subtending an early-branching and highly divergent form of CME, which is conserved and likely functionally important across the kinetoplastid parasites.<\/jats:p>","DOI":"10.1242\/jcs.191478","type":"journal-article","created":{"date-parts":[[2017,2,24]],"date-time":"2017-02-24T01:24:21Z","timestamp":1487899461000},"page":"1379-1392","source":"Crossref","is-referenced-by-count":20,"title":["Lineage-specific proteins essential for endocytosis in trypanosomes"],"prefix":"10.1242","volume":"130","author":[{"given":"Paul T.","family":"Manna","sequence":"first","affiliation":[{"name":"School of Life Sciences, University of Dundee 1 , Dundee, Scotland DD1 5EH , UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Samson O.","family":"Obado","sequence":"additional","affiliation":[{"name":"The Rockefeller University 2 , 1230 York Avenue, New York, NY 10021 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1746-2450","authenticated-orcid":false,"given":"Cordula","family":"Boehm","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Dundee 1 , Dundee, Scotland DD1 5EH , UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catarina","family":"Gadelha","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Nottingham 3 , Nottingham NG2 7UH , UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Andrej","family":"Sali","sequence":"additional","affiliation":[{"name":"California Institute for Quantitative Biosciences, University of California 4 , San Francisco, CA 94158 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian T.","family":"Chait","sequence":"additional","affiliation":[{"name":"The Rockefeller University 2 , 1230 York Avenue, New York, NY 10021 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Michael P.","family":"Rout","sequence":"additional","affiliation":[{"name":"The Rockefeller University 2 , 1230 York Avenue, New York, NY 10021 , USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4817-6035","authenticated-orcid":false,"given":"Mark C.","family":"Field","sequence":"additional","affiliation":[{"name":"School of Life Sciences, University of Dundee 1 , Dundee, Scotland DD1 5EH , UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"237","reference":[{"key":"2024061316415209500_JCS191478C1","doi-asserted-by":"publisher","first-page":"440","DOI":"10.1111\/tra.12040","article-title":"Proteomic analysis of clathrin interactions in trypanosomes reveals dynamic evolution of endocytosis","volume":"14","author":"Adung'a","year":"2013","journal-title":"Traffic"},{"key":"2024061316415209500_JCS191478C2","doi-asserted-by":"publisher","first-page":"75","DOI":"10.1016\/j.molbiopara.2004.10.002","article-title":"A doubly inducible system for RNA interference and rapid RNAi plasmid construction in Trypanosoma brucei","volume":"139","author":"Alibu","year":"2005","journal-title":"Mol. 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