A Global Analysis of SNX27-retromer Assembly and Cargo Specificity Reveals a Function in Glucose and Metal Ion Transport

Nat Cell Biol. 2013 May;15(5):461-71. doi: 10.1038/ncb2721. Epub 2013 Apr 7.

Abstract

The PDZ-domain-containing sorting nexin 27 (SNX27) promotes recycling of internalized transmembrane proteins from endosomes to the plasma membrane by linking PDZ-dependent cargo recognition to retromer-mediated transport. Here, we employed quantitative proteomics of the SNX27 interactome and quantification of the surface proteome of SNX27- and retromer-suppressed cells to dissect the assembly of the SNX27 complex and provide an unbiased global view of SNX27-mediated sorting. Over 100 cell surface proteins, many of which interact with SNX27, including the glucose transporter GLUT1, the Menkes disease copper transporter ATP7A, various zinc and amino acid transporters, and numerous signalling receptors, require SNX27-retromer to prevent lysosomal degradation and maintain surface levels. Furthermore, we establish that direct interaction of the SNX27 PDZ domain with the retromer subunit VPS26 is necessary and sufficient to prevent lysosomal entry of SNX27 cargo. Our data identify the SNX27-retromer as a major endosomal recycling hub required to maintain cellular nutrient homeostasis.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / genetics
  • Adenosine Triphosphatases / metabolism
  • Blotting, Western
  • Cation Transport Proteins / genetics
  • Cation Transport Proteins / metabolism
  • Computational Biology / methods
  • Copper-Transporting ATPases
  • Culture Media / metabolism
  • Glucose / metabolism*
  • Glucose Transporter Type 1 / genetics
  • Glucose Transporter Type 1 / metabolism
  • HeLa Cells
  • Humans
  • Ion Transport
  • Isotope Labeling / methods
  • Lysosomes / metabolism
  • Multiprotein Complexes / metabolism
  • PDZ Domains
  • Protein Folding
  • Protein Interaction Mapping
  • Protein Transport
  • Proteolysis
  • Proteomics / methods*
  • RNA, Small Interfering / genetics
  • RNA, Small Interfering / metabolism
  • Receptor, Platelet-Derived Growth Factor beta / genetics
  • Receptor, Platelet-Derived Growth Factor beta / metabolism
  • Receptors, TNF-Related Apoptosis-Inducing Ligand / genetics
  • Receptors, TNF-Related Apoptosis-Inducing Ligand / metabolism
  • Sorting Nexins / analysis*
  • Sorting Nexins / genetics
  • Sorting Nexins / metabolism
  • Vesicular Transport Proteins / genetics
  • Vesicular Transport Proteins / metabolism

Substances

  • Cation Transport Proteins
  • Culture Media
  • Glucose Transporter Type 1
  • Multiprotein Complexes
  • RNA, Small Interfering
  • Receptors, TNF-Related Apoptosis-Inducing Ligand
  • SLC2A1 protein, human
  • SNX27 protein, human
  • Sorting Nexins
  • TNFRSF10A protein, human
  • VPS26A protein, human
  • VPS35 protein, human
  • Vesicular Transport Proteins
  • Receptor, Platelet-Derived Growth Factor beta
  • Adenosine Triphosphatases
  • ATP7A protein, human
  • Copper-Transporting ATPases
  • Glucose