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Contact-ID, a new tool for profiling organelle-membrane contact sites, reveals regulatory proteins of mitochondrial-associated membrane formation

ABSTRACT: The mitochondria-associated membrane (MAM) has emerged as a cellular signaling hub regulating various cellular processes. However, its molecular components remain unclear owing to lack of reliable methods to purify the intact MAM proteome in a physiological context. Here, we introduce Contact-ID, a split-pair system of BioID with strong activity, for identification of the MAM proteome in live cells. Contact-ID specifically labeled proteins proximal to the contact sites of the endoplasmic reticulum (ER) and mitochondria, and thereby identified 115 MAM-specific proteins. The identified MAM proteins were largely annotated with the outer mitochondrial membrane (OMM) and ER membrane proteins with MAM-related functions: e.g., FKBP8, an OMM protein, facilitated MAM formation and local calcium transport at the MAM. Furthermore, the definitive identification of biotinylation sites revealed membrane topologies of 85 integral membrane proteins. Contact-ID revealed regulatory proteins for MAMformation and could be reliably utilized to profile the proteome at any organelle–membrane contact sites in live cells.

INSTRUMENT(S): Thermo fusion lumos

ORGANISM(S): Homo Sapiens (ncbitaxon:9606)

SUBMITTER: Sang Ki Park Ji Young Mun Jong-Seo Kim Hyun-Woo Rhee

PROVIDER: MSV000084362 | MassIVE |

SECONDARY ACCESSION(S): PXD015534

REPOSITORIES: MassIVE

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