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Spatial proteomics defines the content of trafficking vesicles captured by golgin tethers

Author

Listed:
  • John J. H. Shin

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

  • Oliver M. Crook

    (University of Cambridge
    University of Cambridge
    MRC Biostatistics Unit, School of Clinical Medicine, University of Cambridge)

  • Alicia C. Borgeaud

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

  • Jérôme Cattin-Ortolá

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

  • Sew Y. Peak-Chew

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

  • Lisa M. Breckels

    (University of Cambridge)

  • Alison K. Gillingham

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

  • Jessica Chadwick

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

  • Kathryn S. Lilley

    (University of Cambridge
    University of Cambridge)

  • Sean Munro

    (MRC Laboratory of Molecular Biology, Francis Crick Avenue)

Abstract

Intracellular traffic between compartments of the secretory and endocytic pathways is mediated by vesicle-based carriers. The proteomes of carriers destined for many organelles are ill-defined because the vesicular intermediates are transient, low-abundance and difficult to purify. Here, we combine vesicle relocalisation with organelle proteomics and Bayesian analysis to define the content of different endosome-derived vesicles destined for the trans-Golgi network (TGN). The golgin coiled-coil proteins golgin-97 and GCC88, shown previously to capture endosome-derived vesicles at the TGN, were individually relocalised to mitochondria and the content of the subsequently re-routed vesicles was determined by organelle proteomics. Our findings reveal 45 integral and 51 peripheral membrane proteins re-routed by golgin-97, evidence for a distinct class of vesicles shared by golgin-97 and GCC88, and various cargoes specific to individual golgins. These results illustrate a general strategy for analysing intracellular sub-proteomes by combining acute cellular re-wiring with high-resolution spatial proteomics.

Suggested Citation

  • John J. H. Shin & Oliver M. Crook & Alicia C. Borgeaud & Jérôme Cattin-Ortolá & Sew Y. Peak-Chew & Lisa M. Breckels & Alison K. Gillingham & Jessica Chadwick & Kathryn S. Lilley & Sean Munro, 2020. "Spatial proteomics defines the content of trafficking vesicles captured by golgin tethers," Nature Communications, Nature, vol. 11(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:11:y:2020:i:1:d:10.1038_s41467-020-19840-4
    DOI: 10.1038/s41467-020-19840-4
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