biostudies-literatureBrock KNational Institutes of HealthNIGMS NIH HHSNIH HHS12-31https://www.ebi.ac.uk/biostudies/studies/S-EPMC11599474biostudies-literatureUnknown82(1-2)Focal adhesions serve as structural and signaling hubs, facilitating bidirectional communication at the cell-extracellular matrix interface. Paxillin and the related Hic-5 (TGFβ1i1) are adaptor/scaffold proteins that recruit numerous structural and regulatory proteins to focal adhesions, where they perform both overlapping and discrete functions. In this study, paxillin and Hic-5 were expressed in U2OS osteosarcoma cells as biotin ligase (BioID2) fusion proteins and used as bait proteins for proximity-dependent biotinylation in order to directly compare their respective interactomes. The fusion proteins localized to both focal adhesions and the centrosome, resulting in biotinylation of components of each of these structures. Biotinylated proteins were purified and analyzed by mass spectromCytoskeleton (Hoboken, N.J.)A comparative analysis of paxillin and Hic-5 proximity interactomes.PMC11599474S10 1S10OD023617‐01A1S10 1S10OD023617-01A1S10 OD023617R35 GM131709Turner CEDe Jong EPAlpha KMBrennan GLuke EBrock KfalseA comparative analysis of paxillin and Hic-5 proximity interactomes.Focal adhesions serve as structural and signaling hubs, facilitating bidirectional communication at the cell-extracellular matrix interface. Paxillin and the related Hic-5 (TGFβ1i1) are adaptor/scaffold proteins that recruit numerous structural and regulatory proteins to focal adhesions, where they perform both overlapping and discrete functions. In this study, paxillin and Hic-5 were expressed in U2OS osteosarcoma cells as biotin ligase (BioID2) fusion proteins and used as bait proteins for proximity-dependent biotinylation in order to directly compare their respective interactomes. The fusion proteins localized to both focal adhesions and the centrosome, resulting in biotinylation of components of each of these structures. Biotinylated proteins were purified and analyzed by mass spectrom2025-01-01T00:00:00Z2025 Jan2026-06-06T09:03:02.791Z2026-05-28T03:10:42.037ZS-EPMC115994743880109810.1002/cm.21878