Dataset Information

GIPC3 AP-MS/MS from chick inner ear extracts

ABSTRACT: To understand how GIPC3 exerts its effects on cuticular plate dimensions, we examined the GIPC3 protein-interaction network in hair cells. We immunoaffinity purified GIPC3 from crosslinked chicken inner ear extracts that were enriched for stereocilia, but still contain large amounts of hair-cell cytoplasmic proteins (Morgan et al., 2016). We carried out two separate experiments, each with ~1000 chicken ears, where we stabilized protein complexes using primary amine-reactive homo-bifunctional N-hydroxysuccimide ester crosslinkers that are thiol-cleavable and hence reversible (Mattson et al., 1993). In one experiment, we used dithiobis(succinimidyl propionate) (DSP), a membrane-permeable crosslinker that crosslinks extracellular and intracellular complexes; in the other experiment, we used 3,3'-dithiobis(sulfosuccinimidyl propionate) (DTSSP), which is membrane impermeant and thus only stabilizes extracellular and transmembrane complexes. We prepared soluble extracts of crude, crosslinked stereocilia and used these fractions for identifying GIPC3-interacting proteins.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Gallus Gallus (chicken)

TISSUE(S): Ear Hair Cell, Inner Ear

SUBMITTER: Peter Barr-Gillespie

LAB HEAD: Peter Barr-Gillespie

PROVIDER: PXD038234 | Pride | 2023-05-09

REPOSITORIES: Pride

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Publications

GIPC3 couples to MYO6 and PDZ domain proteins, and shapes the hair cell apical region.

Chatterjee Paroma P Morgan Clive P CP Krey Jocelyn F JF Benson Connor C Goldsmith Jennifer J Bateschell Michael M Ricci Anthony J AJ Barr-Gillespie Peter G PG

Journal of cell science 20230525 10

GIPC3 has been implicated in auditory function. Here, we establish that GIPC3 is initially localized to the cytoplasm of inner and outer hair cells of the cochlea and then is increasingly concentrated in cuticular plates and at cell junctions during postnatal development. Early postnatal Gipc3KO/KO mice had mostly normal mechanotransduction currents, but had no auditory brainstem response at 1 month of age. Cuticular plates of Gipc3KO/KO hair cells did not flatten during development as did those ...[more]

PMID: 37096733

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Dataset Information

GIPC3 AP-MS/MS from chick inner ear extracts

Publications

GIPC3 couples to MYO6 and PDZ domain proteins, and shapes the hair cell apical region.

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