Mapping and Quantitation of the Interaction Between the Recombination Activating Gene Proteins RAG1 and RAG2

Zhang Y, et al., 290(19):11802-17, J Biol Chem, 2015

Described herein is an investigation of interaction between the Recombination Activating Gene (RAG) proteins, RAG1 and RAG2 using Bio-Layer Interferometry (BLI) and pulldown assays. A Pall ForteBio BLItz instrument was used to measure the RAG1-RAG2 binding affinity. In these experiments, GST-R2c and MBP-R1c-His6 were immobilized on to Anti-GST or Anti-His Biosensor probes, respectively. Overall results suggests that the RAG1-RAG2 interaction requires only a small portion of RAG1 flanking the RAG1 catalytic region. The binding affinity (KD) for the RAG1-RAG2 interaction is around 0.4 μM.

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