Protein-protein interactions govern almost all cellular functions. These complex networks of stable and transient associations can be mapped by affinity purification mass spectrometry (AP-MS) and complementary proximity-based labeling methods such as BioID. To exploit the advantages of both strategies, we here design and optimize an integrated approach combining AP-MS and BioID in a single construct, which we term MAC-tag. We systematically apply the MAC-tag approach to 18 subcellular and 3 sub-organelle localization markers, generating a molecular context database, which can be used to define a protein's molecular location. In addition, we show that combining the AP-MS and BioID results makes it possible to obtain interaction distances within a protein complex. Taken together, our integrated strategy enables the comprehensive mapping of the physical and functional interactions of proteins, defining their molecular context and improving our understanding of the cellular interactome.

中文翻译：

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兼容AP-MS和BioID的MAC标签可对蛋白质相互作用和亚细胞定位进行全面定位。

蛋白质-蛋白质相互作用控制着几乎所有的细胞功能。可以通过亲和纯化质谱（AP-MS）和基于互补性的邻近标记方法（例如BioID）来映射这些稳定且瞬态关联的复杂网络。为了利用这两种策略的优势，我们在这里设计并优化了在单个结构中将AP-MS和BioID结合在一起的集成方法，我们将其称为MAC标签。我们系统地将MAC标签方法应用于18个亚细胞和3个亚细胞器的定位标记，生成分子上下文数据库，该数据库可用于定义蛋白质的分子位置。此外，我们表明结合AP-MS和BioID结果可以在蛋白质复合物中获得相互作用距离。在一起