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In situ cell-surface proteomic profiling in mice

ABSTRACT: Shuster SA, Li J, Chon U, Hu MC, Luginbuhl DJ, Udeshi ND, Carey DK, Takeo YH, Xu C, Mani D.R., Han S, Ting AY, Carr SA, Luo L. Cell-surface proteins mediate cell-cell interactions throughout the lifetime of multicellular organisms, yet there are no general methods for profiling them in vertebrate tissues. Here, we present in-situ cell-surface proteome extraction by extracellular labeling (iPEEL) in mice, which enables cell-type-specific profiling of cell-surface proteomes in native tissues. Applying iPEEL to developing and mature cerebellar Purkinje cells revealed a differential enrichment in cell-surface proteins with post-translational processing and synaptic functions in the developing and mature proteomes, respectively. A proteome-instructed in vivo loss-of-function screen identified a critical role for Armadillo-like helical domain-containing protein 4 (Armh4) in Purkinje cell dendrite morphogenesis. Armh4 overexpression also disrupts dendrite morphogenesis; this effect requires its conserved cytoplasmic domain and is augmented by disruption of endocytosis. Our results highlight the utility of cell-surface proteomic profiling in native tissues in identifying regulators of cell-surface signaling events.

INSTRUMENT(S): Q Exactive HF-X, Q Exactive Plus

ORGANISM(S): Mus Musculus (ncbitaxon:10090)

SUBMITTER: Steven A. Carr

PROVIDER: MSV000088506 | MassIVE | Fri Dec 03 12:41:00 GMT 2021

REPOSITORIES: MassIVE

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