Skip to content

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

Research output: Contribution to journalArticle

Original languageEnglish
Pages (from-to)1670-1676
Number of pages7
JournalCurrent Biology
Volume24
Issue number14
Early online date21 Jul 2014
DOIs
DateAccepted/In press - 10 Jun 2014
DateE-pub ahead of print - 21 Jul 2014
DatePublished (current) - 21 Jul 2014

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.

    Research areas

  • Ankyrin Repeat, Antigens, Neoplasm, Binding Sites, Cell Line, Tumor, Cells, Cultured, Endosomes, Golgi Apparatus, HeLa Cells, Humans, Microfilament Proteins, Molecular Chaperones, Mutation, Parkinson Disease, Protein Binding, Protein Transport, Vesicular Transport Proteins, Journal Article, Research Support, N.I.H., Extramural, Research Support, Non-U.S. Gov't

Download statistics

No data available

Documents

Documents

  • Full-text PDF (final published version)

    Rights statement: This is the final published version of the article (version of record). It first appeared online via Elsevier (Cell Press) at http://www.sciencedirect.com/science/article/pii/S0960982214007404. Please refer to any applicable terms of use of the publisher

    Final published version, 2.18 MB, PDF document

    Licence: CC BY

DOI

View research connections

Related faculties, schools or groups