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VHS domains of ESCRT-0 cooperate in high-avidity binding to polyubiquitinated cargo.

The EMBO journal | Mar 17, 2010

VHS (Vps27, Hrs, and STAM) domains occur in ESCRT-0 subunits Hrs and STAM, GGA adapters, and other trafficking proteins. The structure of the STAM VHS domain-ubiquitin complex was solved at 2.6 A resolution, revealing that determinants for ubiquitin recognition are conserved in nearly all VHS domains. VHS domains from all classes of VHS-domain containing proteins in yeast and humans, including both subunits of ESCRT-0, bound ubiquitin in vitro. ESCRTs have been implicated in the sorting of Lys63-linked polyubiquitinated cargo. Intact human ESCRT-0 binds Lys63-linked tetraubiquitin 50-fold more tightly than monoubiquitin, though only 2-fold more tightly than Lys48-linked tetraubiquitin. The gain in affinity is attributed to the cooperation of flexibly connected VHS and UIM motifs of ESCRT-0 in avid binding to the polyubiquitin chain. Mutational analysis of all the five ubiquitin-binding sites in yeast ESCRT-0 shows that cooperation between them is required for the sorting of the Lys63-linked polyubiquitinated cargo Cps1 to the vacuole.

Pubmed ID: 20150893 RIS Download

Mesh terms: Amino Acid Motifs | Binding Sites | Crystallography, X-Ray | Endosomal Sorting Complexes Required for Transport | Humans | Molecular Sequence Data | Polyubiquitin | Protein Structure, Tertiary | Saccharomyces cerevisiae | Saccharomyces cerevisiae Proteins | Ubiquitination

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Associated grants

  • Agency: Intramural NIH HHS, Id: Z01 DK036126-02

Addgene (Reagent, Plasmid)

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