Protocol to differentially quantify spatially resolved viral protein-cellular protein interactions via proximity
Helene Hoenigsperger1, Susanne Klute1, Zoé Engels1
1Institute of Molecular Virology, Ulm University Medical Center, 89081 Ulm, Baden-Wuerttemberg, Germany.
None:
The spatial organization of viral and cellular proteins shapes signal transduction. Here, we present a protocol to quantify differential protein-protein interactions by measuring their spatial association using proximity ligation assays (PLAs). We describe steps for seeding, transfection, proximity labeling, and confocal microscopy imaging and provide procedures for quantitative analysis. Our approach complements co-immunoprecipitation-based interactome data by enabling in situ quantification of differential binding between cellular and viral interaction partners. For complete details on the use and execution of this protocol, please refer to Klute et al.1.
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