title: Illuminating allostery in metal sensing transcriptional regulators.
abstract: The intracellular availability of all biologically required transition metal ions in bacteria, e.g., Zn, Cu, Fe, as well as the detoxification of nonbiological heavy metal pollutants, is controlled at the molecular level by a panel of metalloregulatory or "metal sensor" proteins. Metal sensor proteins are specialized allosteric proteins that regulate the transcription of genes linked to transition metal homeostasis as a result of direct binding of a single metal ion or two closely related metal ions, to the exclusion of all others. In many cases, the binding of the cognate metal ion induces a structural change in a metal sensor oligomer that either activates or inhibits operator DNA binding. A quantitative measure of the degree to which a particular metal drives metalloregulation of transcription is the allosteric coupling-free energy, ΔG(c). In this chapter, we outline detailed spectroscopically derived methods for measuring metal binding affinity, K(Me), as well as ΔG(c) independent of K(Me), presented in the context of a simple coupled equilibrium scheme. Studies carried out in this way provide quantitative insights into the degree to which a particular metal ion is capable of driving allosteric switching, and via ligand substitution, the extent to which individual coordination bonds establish structural linkage of allosteric metal and operator DNA-binding sites.
authors.affiliation: Winthrop University, Rock Hill, SC, USA. grossoehmen@winthrop.edu
authors.forename: Nicholas E
authors.initials: NE
authors.lastname: Grossoehme
authors.forename: David P
authors.initials: DP
authors.lastname: Giedroc
mesh_terms: D000494:Allosteric Regulation; D002021:Buffers; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D008667:Metalloproteins / Q000737:chemistry / Q000378:metabolism*; D008670:Metals / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D011485:Protein Binding; D011487:Protein Conformation; D012996:Solutions / Q000737:chemistry; D014157:Transcription Factors / Q000737:chemistry / Q000378:metabolism*
chemical_list: D002021:Buffers; D008667:Metalloproteins; D008670:Metals; D012996:Solutions; D014157:Transcription Factors; D004247:DNA