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000470981 1001_ $$0P:(DE-H253)PIP1086337$$aLizarrondo, Javier$$b0
000470981 245__ $$aStructure of the endocytic adaptor complex reveals the basis for efficient membrane anchoring during clathrin-mediated endocytosis
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000470981 520__ $$aDuring clathrin-mediated endocytosis, a complex and dynamic network of protein-membrane interactions cooperate to achieve membrane invagination. Throughout this process in yeast, endocytic coat adaptors, Sla2 and Ent1, must remain attached to the plasma membrane to transmit force from the actin cytoskeleton required for successful membrane invagination. Here, we present a cryo-EM structure of a 16-mer complex of the ANTH and ENTH membrane-binding domains from Sla2 and Ent1 bound to PIP$_2$ that constitutes the anchor to the plasma membrane. Detailed in vitro and in vivo mutagenesis of the complex interfaces delineate the key interactions for complex formation and deficient cell growth phenotypes demonstrate its biological relevance. A hetero-tetrameric unit binds PIP$_2$ molecules at the ANTH-ENTH interfaces and can form larger assemblies to contribute to membrane remodeling. Finally, a time-resolved small-angle X-ray scattering study of the interaction of these adaptor domains in vitro suggests that ANTH and ENTH domains have evolved to achieve a fast subsecond timescale assembly in the presence of PIP$_2$ and do not require further proteins to form a stable complex. Together, these findings provide a molecular understanding of an essential piece in the molecular puzzle of clathrin-coated endocytic sites.
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000470981 7001_ $$0P:(DE-HGF)0$$aKlebl, David P.$$b1
000470981 7001_ $$0P:(DE-H253)PIP1030856$$aNiebling, Stephan$$b2
000470981 7001_ $$0P:(DE-HGF)0$$aAbella, Marc$$b3
000470981 7001_ $$0P:(DE-HGF)0$$aSchroer, Martin A.$$b4
000470981 7001_ $$0P:(DE-H253)PIP1020991$$aMertens, Haydyn D. T.$$b5
000470981 7001_ $$0P:(DE-H253)PIP1025998$$aVeith, Katharina$$b6
000470981 7001_ $$0P:(DE-H253)PIP1087856$$aThuenauer, Roland$$b7
000470981 7001_ $$0P:(DE-H253)PIP1001422$$aSvergun, Dmitri I.$$b8
000470981 7001_ $$00000-0003-4286-7684$$aSkruzny, Michal$$b9
000470981 7001_ $$00000-0001-9029-1865$$aSobott, Frank$$b10
000470981 7001_ $$00000-0001-6869-4414$$aMuench, Stephen P.$$b11
000470981 7001_ $$0P:(DE-HGF)0$$aGarcia-Alai, Maria M.$$b12$$eCorresponding author
000470981 773__ $$0PERI:(DE-600)2553671-0$$a10.1038/s41467-021-23151-7$$gVol. 12, no. 1, p. 2889$$n1$$p2889 (1-15)$$tNature Communications$$v12$$x2041-1723$$y2021
000470981 8564_ $$uhttps://www.nature.com/articles/s41467-021-23151-7
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