Probing Pairwise Protein-Protein Interactions with the Bacterial POLAR Assay.

Tesseur, Coralie;Laloux, Géraldine
(2025) Proximity-Dependent Protein Biotinylation : Methods and Protocols — ISBN: [978-1-0716-4694-6], p. 103-113, published

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Abstract
The PopZ-linked apical recruitment (POLAR) assay is an in vivo two-hybrid assay performed in Escherichia coli, using epifluorescence microscopy to visualize the recruitment of a prey protein fusion by a bait protein fusion tethered artificially at the cell pole. Here, we present a standardized protocol of the POLAR assay, specifically designed to investigate protein-protein interactions (PPIs) in the cytoplasm. Notably, the POLAR assay has been successfully used to validate potential PPIs identified through proximity labeling, highlighting its relevance as a complementary tool within the broader context of studying protein interaction networks.
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Tesseur, C., & Laloux, G. (2025). Probing Pairwise Protein-Protein Interactions with the Bacterial POLAR Assay. In Petra Van Damme (ed.) (ed.), Proximity-Dependent Protein Biotinylation : Methods and Protocols (p. p. 103-113). Humana. https://doi.org/10.1007/978-1-0716-4694-6_7