PNNL

Abstract

The rapid isolation of protein complexes is critical to the goal of interpreting genomic information in the context of cellular function. High-throughput methods for identifying protein binding partners in a way suitable for mass spectrometric identification and structural analysis are sought after and small molecule/peptide interactions may provide the key. We have now shown that a newly synthesized resin derivatized with a bisarsenical dye can be used to isolate the Shewanella oneidensis RNA polymerase core enzyme, using the tetracysteine-tagged RNA polymerase A as bait protein. A critical advantage of this method is the ability to release the intact complex using a mild, one-step procedure with a competing dithiol. In addition to the identification of the core subunit complex, potential additional regulatory factors, including universal stress protein, were identified. These results provide a path forward to identifying how changes in critical protein complexes over time modulate cell function