Proximity-based proteomic profiling uncovers distinct interactome of human RAG1 and RAG2

• TurboID-based proximity labeling identified 88 RAG1- and 146 RAG2-associated proteins, with only 23 shared, revealing distinct interactomes. • RAG1 and RAG2 exhibit subunit-specific interaction networks, suggesting independent regulatory roles beyond the heterotetramer. • The human RAG interactome includes proteins involved in nuclear transport, chromatin remodeling, and DNA repair, expanding known RAG functions. • This study provides a comprehensive framework for understanding RAG's multifaceted roles in adaptive immunity and potential disease associations.