Functional characterization of a purified, recombinant NF-kappaB/IkappaB complex.

Abstract

The NF-kappaB signal transduction pathway involves the interaction of several NF-kappaB and IkappaB family members that are activated by a diverse range of extracellular signals and that stimulate many different cellular responses. The biochemical regulation of this cascade can be studied by establishing a cell-free system using purified proteins. As a first step toward establishing an in vitro model incorporating multiple combinations of NF-kappaB and IkappaB proteins, we produced purified human p65 (RelA) and human IkappaBalpha proteins and tested their functionality. Full-length RelA and IkappaBalpha proteins were overproduced by coinfection of TN5-JE cells with two recombinant baculoviruses. RelA and IkappaBalpha formed a stable complex that could be purified to >95% homogeneity. Protein-protein interactions, protein-DNA binding, and protein phosphorylation were similar to the native proteins.

Cite this paper

@article{Larson1999FunctionalCO, title={Functional characterization of a purified, recombinant NF-kappaB/IkappaB complex.}, author={Chris J Larson and Ying Wei Li and Frank Mercurio and Rolf M{\"{u}ller}, journal={Biochemical and biophysical research communications}, year={1999}, volume={260 3}, pages={691-8} }