Protein-protein interaction between an RNA polymerase sigma factor sR and its corresponding zinc binding anti-sigma (ZAS) protein RsrA triggers thioredoxin system in S. coelicolor. RsrA with zinc ion associates to sR repressing its transcriptional activity in reduced environment of cytoplasm, while during disulfide stress it forms intramolecular disulfide bonds, releases zinc, and dissociates sR. We designed a pair of FRET-based probes for real-time monitoring protein-protein interaction between sR and RsrA, and how this anti-oxidative stress system initiates in response to oxidants. The FRET-based method also confirms the function of zinc coordination and conformational change induced by ROS in RsrA, clarifying the difference between Zn-RsrA and apo-RsrA. Sensitivity of the redox switch to various ROS deriving from different metabolite pathways was determined and kinetic constants within the interaction were obtained by the FRET-based method. Characterizations of the probes were also verified by Octet analysis and far-UV circular dichroism.

FRET-based method for probing protein-protein interaction between [sigma]<sup>R </sup>and RsrA from <i>Streptomyces coelicolor</i> in response to redox environment in real time

245th National Meeting (2013)

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FRET-based method for probing protein-protein interaction between [sigma]R and RsrA from Streptomyces coelicolor in response to redox environment in real time

FRET-based method for probing protein-protein interaction between [sigma]^Rand RsrA from Streptomyces coelicolor in response to redox environment in real time