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

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 and Sean Munro ()
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John J. H. Shin: MRC Laboratory of Molecular Biology, Francis Crick Avenue
Oliver M. Crook: 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
Sean Munro: MRC Laboratory of Molecular Biology, Francis Crick Avenue

Nature Communications, 2020, vol. 11, issue 1, 1-13

Abstract: 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.

Date: 2020
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DOI: 10.1038/s41467-020-19840-4

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