Large Scale Production of Nuclear Receptor Ligand-Binding Domains

AbstractThe nuclear receptor superfamily is composed of over 150 inducible transcription factors, most of which (1-2) do not have well characterized ligands to date. Nuclear receptors regulate promoters through specific protein-protein and protein-DNA interaction at their hormone response elements, and up-or downregulate their target genes in a ligand-dependent manner with the aid of various cofactors (2-4). The high-affinity binding of ligands sets into action the complex signal transduction properties of these proteins. Ligand occupancy is a key determinant of function, because it can result in the formation of distinct molecular surfaces that have enhanced affinity for coactivators or corepressors (5-7). However, additional levels of regulation can be achieved through interactions with other systems, such as molecular chaperones, cyclic adenosine monophosphate-regulated kinases and the AP-1 activator (8-9). All of these interactions together form a rich and elaborate network by which the ligands ultimately exert their powerful effects on gene expression.KeywordsNuclear ReceptorRefold BufferHormone Response ElementInsoluble PortionFreeze Cell PelletThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.