PT  - JOURNAL ARTICLE
AU  - Harpreet Kaur Salgania
AU  - Jutta Metz
AU  - Mandy Jeske
TI  - ReLo: a simple colocalization assay to identify and characterize physical protein-protein interactions
AID  - 10.1101/2022.03.04.482790
DP  - 2022 Jan 01
TA  - bioRxiv
PG  - 2022.03.04.482790
4099  - http://biorxiv.org/content/early/2022/03/04/2022.03.04.482790.short
4100  - http://biorxiv.org/content/early/2022/03/04/2022.03.04.482790.full
AB  - Characterizing protein-protein interactions (PPIs) is fundamental for understanding biochemical processes. Many methods have been established to identify and study direct PPIs; however, screening and investigating PPIs involving large and poorly soluble proteins remain challenges. As a result, we developed ReLo, a simple cell culture-based method to detect and investigate interactions in a cellular context. ReLo allows both the identification of protein domains that mediate the formation of complexes and screening of interfering point mutations. ReLo is sensitive to drugs that mediate or interfere with a specific interaction. Furthermore, protein conformation- and protein arginine methylation-dependent interactions can be studied. Importantly, ReLo can be used for specifically detecting direct but not indirect PPIs and can be applied for describing the binding topology of subunits within multiprotein complexes. Because of these attributes, ReLo is a simple, quick and versatile tool for identifying and studying binary PPIs, as well as for characterizing multisubunit complexes.Competing Interest StatementThe authors have declared no competing interest.