ArticleBio-protocol2026
Identification of the Subcompartment-Specific Mitochondrial Proteome by APEX2 Proximity Labeling in
Article in Bio-protocol, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
The cellular compartments of eukaryotic cells are defined by their specific protein compositions. Different strategies are used for the identification of the subcellular proteomes, such as fractionation by differential centrifugation of cellular extracts. The localization of mitochondrial proteins is particularly challenging, as mitochondria consist of two membranes of different protein composition and two aqueous subcompartments, the intermembrane space (IMS) and the matrix. Previous studies identified subcompartment-specific proteomes by using combinations of hypotonic swelling and protease digestion followed by mass spectrometry. Here, we present an alternative, more unbiased method to identify the proteomes of mitochondrial subcompartments by use of an improved ascorbate peroxidase (APEX2) that is targeted to the IMS and the matrix. This method allows the subcompartment-specific labeling of proteins in mitochondria isolated from cells of the baker's yeast
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