Revealing the Affinity of Individual and Combined Flig Domains for Flim in the Bacterial Flagellar Motor Switch Complex by CG-MALS

Abstract

measurements, stopped-flow fluorescence and ITC. We are using pyrene labeled DNA, IkBa, or NF-kb to study the fluorescence changes occurring during the enhanced dissociation process. Our results show that IkBa increases the dissociation rate of the DNA from the NF-kB complex in a concentration-dependent manner and with high efficiency. We also repeated the experiments using different DNA chains and different mutants of IkBa. We are also studying IkBb, which appears to stabilize the NF-kB/DNA interaction. Both IkBa and IkBb are able to form NF-kB/DNA/IkB ternary complexes. The rates of association and dissociation of DNA to form the ternary complexes was also measured and compared to interpret the kinetics of the enhanced dissociation process.