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Proximity-Labeling MS (APEX-MS) | Krogan Lab

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For this approach the protein of interest is fused to a peroxidase (ascorbic acid peroxidase; APEX). Upon treatment with H2O2, APEX converts exogenously supplied biotin-phenol to biotin-phenoxyl radicals, which results in a covalent labeling of protein in a radius of 20nm. Subsequently, the biotinylated proteins are enriched using Steptavidin. This technology allows for the first time to take ‘snapshots’ of local protein environments at a given time and to capture transient protein interactions. Proximity-based labeling approaches have advanced rapidly in the last several years and have been shown to provide complementary results to traditional affinity-based purification methods (AP-MS), thus having the potential to open entirely new avenues of research and provide novel insights into protein functions. APEX has been used to capture entire organelle proteomes with high temporal resolution, but its breadth of labeling is generally thought to preclude the higher spatial resolution neces

For this approach the protein of interest is fused to a peroxidase (ascorbic acid peroxidase; APEX ). Upon treatment with H 2 O 2 , APEX converts exogenously supplied biotin-phenol to biotin-phenoxyl radicals, which results in a covalent labeling of protein in a radius of 20nm. Subsequently, the biotinylated proteins are enriched using Steptavidin. This technology allows for the first time to take ‘snapshots’ of local protein environments at a given time and to capture transient protein interactions. Proximity-based labeling approaches have advanced rapidly in the last several years and have b

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