2. Academic Validation
  3. Retromer binding to FAM21 and the WASH complex is perturbed by the Parkinson disease-linked VPS35(D620N) mutation

Retromer binding to FAM21 and the WASH complex is perturbed by the Parkinson disease-linked VPS35(D620N) mutation

  • Curr Biol. 2014 Jul 21;24(14):1670-1676. doi: 10.1016/j.cub.2014.06.024.
Ian J McGough 1,  Florian Steinberg 1,  Da Jia 2,  Peter A Barbuti 3,  Kirsty J McMillan 1,  Kate J Heesom 4,  Alan L Whone 5,  Maeve A Caldwell 3,  Daniel D Billadeau 6,  Michael K Rosen 7,  Peter J Cullen 8
Affiliations

Affiliations

  • 1 The Henry Wellcome Integrated Signaling Laboratories, School of Biochemistry, Medical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
  • 2 Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
  • 3 Henry Wellcome Laboratory for Integrative Neuroscience and Endocrinology, School of Clinical Sciences, University of Bristol, Dorothy Hodgkin Building, Whitson Street, Bristol BS1 3NY, UK.
  • 4 Proteomics Facility, School of Biochemistry, Medical Sciences Building, University of Bristol, Bristol BS8 1TD, UK.
  • 5 Institute of Clinical Neurosciences, University of Bristol, Frenchay Hospital, Bristol BS16 1LE, UK.
  • 6 Departments of Biochemistry and Molecular Biology and Immunology, Mayo Clinic, Rochester, MN 55905, USA.
  • 7 Department of Biophysics, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA; Howard Hughes Medical Institute, University of Texas Southwestern Medical Center, Dallas, TX 75390, USA.
  • 8 The Henry Wellcome Integrated Signaling Laboratories, School of Biochemistry, Medical Sciences Building, University of Bristol, Bristol BS8 1TD, UK. Electronic address: pete.cullen@bristol.ac.uk.
PMID: 24980502 DOI: 10.1016/j.cub.2014.06.024
Abstract

Retromer is a protein assembly that plays a central role in orchestrating export of transmembrane-spanning cargo proteins from endosomes into retrieval pathways destined for the Golgi apparatus and the plasma membrane [1]. Recently, a specific mutation in the retromer component VPS35, VPS35(D620N), has linked retromer dysfunction to familial autosomal dominant and sporadic Parkinson disease [2, 3]. However, the effect of this mutation on retromer function remains poorly characterized. Here we established that in cells expressing VPS35(D620N) there is a perturbation in endosome-to-TGN transport but not endosome-to-plasma membrane recycling, which we confirm in patient cells harboring the VPS35(D620N) mutation. Through comparative stable isotope labeling by Amino acids in Cell Culture (SILAC)-based analysis of wild-type VPS35 versus the VPS35(D620N) mutant interactomes, we establish that the major defect of the D620N mutation lies in the association to the actin-nucleating Wiskott-Aldrich syndrome and SCAR homolog (WASH) complex. Moreover, using isothermal calorimetry, we establish that the primary defect of the VPS35(D620N) mutant is a 2.2 ± 0.5-fold decrease in affinity for the WASH complex component FAM21. These data define the primary molecular defect in retromer assembly that arises from the VPS35(D620N) mutation and, by revealing functional effects on retromer-mediated endosome-to-TGN transport, provide new insight into retromer deregulation in Parkinson disease.

Figures