Eukaryotic cells rely on endocytosis to regulate their plasma membrane proteome and lipidome. Most eukaryotic groups, except fungi and animals, have retained the evolutionary ancient TSET complex as an endocytic regulator. Unlike other coatomer complexes, structural insight into TSET is lacking. Here, we reveal the molecular architecture of plant TSET [TPLATE complex (TPC)] using an integrative structural approach. We identify crucial roles for specific TSET subunits in complex assembly and membrane interaction. Our data therefore generate fresh insight into the differences between the hexameric TSET in Dictyostelium and the octameric TPC in plants. Structural elucidation of this ancient adaptor complex represents the missing piece in the coatomer puzzle and vastly advances our functional as well as evolutionary insight into the process of endocytosis.

Department of Biochemistry and Microbiology Ghent University 9052 Ghent Belgium

Department of Plant Biotechnology and Bioinformatics Ghent University Technologiepark 71 9052 Ghent Belgium

Institute of Experimental Botany Academy of Sciences of the Czech Republic Rozvojová 263 16502 Prague 6 Czech Republic

VIB Center for Inflammation Research 9052 Ghent Belgium

VIB Center for Plant Systems Biology Technologiepark 71 9052 Ghent Belgium

VIB VUB Center for Structural Biology Structural Biology Research Center Molecular Recognition Unit 1050 Brussels Belgium

Vrije Universiteit Brussel Structural Biology Brussels Department of Biotechnology 1050 Brussels Belgium

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Biomolecular condensation orchestrates clathrin-mediated endocytosis in plants

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Distinct EH domains of the endocytic TPLATE complex confer lipid and protein binding