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-Structure paper
タイトル | Improved mammalian retromer cryo-EM structures reveal a new assembly interface. |
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ジャーナル・号・ページ | J Biol Chem, Vol. 298, Issue 11, Page 102523, Year 2022 |
掲載日 | 2022年9月26日 |
著者 | Amy K Kendall / Mintu Chandra / Boyang Xie / William Wan / Lauren P Jackson / |
PubMed 要旨 | Retromer (VPS26/VPS35/VPS29 subunits) assembles with multiple sorting nexin proteins on membranes to mediate endosomal recycling of transmembrane protein cargoes. Retromer has been implicated in ...Retromer (VPS26/VPS35/VPS29 subunits) assembles with multiple sorting nexin proteins on membranes to mediate endosomal recycling of transmembrane protein cargoes. Retromer has been implicated in other cellular processes, including mitochondrial homeostasis, nutrient sensing, autophagy, and fission events. Mechanisms for mammalian retromer assembly remain undefined, and retromer engages multiple sorting nexin proteins to sort cargoes to different destinations. Published structures demonstrate mammalian retromer forms oligomers in vitro, but several structures were poorly resolved. We report here improved retromer oligomer structures using single-particle cryo-EM by combining data collected from tilted specimens with multiple advancements in data processing, including using a 3D starting model for enhanced automated particle picking in RELION. We used a retromer mutant (3KE retromer) that breaks VPS35-mediated interfaces to determine a structure of a new assembly interface formed by the VPS26A and VPS35 N-termini. The interface reveals how an N-terminal VPS26A arrestin saddle can link retromer chains by engaging a neighboring VPS35 N- terminus, on the opposite side from the well-characterized C-VPS26/N-VPS35 interaction observed within heterotrimers. The new interaction interface exhibits substantial buried surface area (∼7000 Å) and further suggests that metazoan retromer may serve as an adaptable scaffold. |
リンク | J Biol Chem / PubMed:36174678 / PubMed Central |
手法 | EM (単粒子) |
解像度 | 4.5 - 7.3 Å |
構造データ | EMDB-26340: Mouse retromer (VPS26/VPS35 E615A/D616A/E617A mutant/VPS29) 3KE particle sub-structure EMDB-26341: Mouse retromer (VPS26/VPS35 E615A/D616A/E617A mutant/VPS29) 3KE particle EMDB-26342: Mouse retromer (VPS26/VPS35/VPS29) dimer of heterotrimers EMDB-26343: Mouse retromer sub-structure: VPS35/VPS35 curved dimer EMDB-26345: Mouse retromer sub-structure: VPS35/VPS35 flat substructure PDB-7u6f: |
化合物 | ChemComp-GOL: ChemComp-SO4: ChemComp-EDO: ChemComp-HOH: |
由来 |
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キーワード | TRANSPORT PROTEIN / retromer / membrane trafficking / endosomal trafficking / membrane coat complexes / PROTEIN TRANSPORT |