title: Domain stability in the AAA+ ATPase ClpB from Escherichia coli.
abstract: ClpB is a heat-shock protein that reactivates aggregated proteins in cooperation with the DnaK chaperone system. ClpB belongs to the family of AAA+ ATPases and forms ring-shaped oligomers: heptamers in the absence of nucleotides and hexamers in the presence of nucleotides. We investigated the thermodynamic stability of ClpB in its monomeric and oligomeric forms. ClpB contains six distinct structural domains: the N-terminal domain involved in substrate binding, two AAA+ ATP-binding modules, each consisting of two domains, and a coiled-coil domain inserted between the AAA+ modules. We produced seven variants of ClpB, each containing a single Trp located in each of the ClpB domains and measured the changes in Trp fluorescence during the equilibrium urea-induced unfolding of ClpB. We found that two structural domains: the small domain of the C-terminal AAA+ module and the coiled-coil domain were destabilized in the oligomeric form of ClpB, which indicates that only those domains change their conformation and/or interactions during formation of the ClpB rings.
authors.affiliation: Department of Biochemistry, Kansas State University, Manhattan, KS 66506, USA.
authors.forename: Maria
authors.initials: M
authors.lastname: Nagy
authors.forename: Vladimir
authors.initials: V
authors.lastname: Akoev
authors.forename: Michal
authors.initials: M
authors.lastname: Zolkiewski
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000648:ultrastructure; D003198:Computer Simulation; D049071:Endopeptidase Clp; D004795:Enzyme Stability; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000648:ultrastructure; D006360:Heat-Shock Proteins / Q000737:chemistry* / Q000648:ultrastructure; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D014508:Urea / Q000737:chemistry*
chemical_list: D029968:Escherichia coli Proteins; D006360:Heat-Shock Proteins; D014508:Urea; D049071:Endopeptidase Clp; D000251:Adenosine Triphosphatases; C069775:ClpB protein, E coli