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An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping of protein interactions and subcellular localizations

Xiaonan Liu, Kari Salokas, Fitsum Tamene, Yaming Jiu, Rigbe G. Weldatsadik, Tiina Öhman and Markku Varjosalo ()
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Xiaonan Liu: University of Helsinki
Kari Salokas: University of Helsinki
Fitsum Tamene: University of Helsinki
Yaming Jiu: University of Helsinki
Rigbe G. Weldatsadik: University of Helsinki
Tiina Öhman: University of Helsinki
Markku Varjosalo: University of Helsinki

Nature Communications, 2018, vol. 9, issue 1, 1-16

Abstract: Abstract 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.

Date: 2018
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DOI: 10.1038/s41467-018-03523-2

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