title: The chaperoning activity of hsp110. Identification of functional domains by use of targeted deletions.
abstract: hsp110 is one of major heat shock proteins of eukaryotic cells and is a diverged relative of the hsp70 family. It has been previously shown that hsp110 maintains heat-denatured luciferase in a soluble, folding competent state and also confers cellular heat resistance in vivo. In the present study the functional domains of hsp110 that are responsible for its chaperoning activity are identified by targeted deletion mutagenesis using the DnaK structure as the model. The chaperoning activity of mutants is assessed based on their ability to solubilize heat-denatured luciferase as well as to refold luciferase in the presence of rabbit reticulocyte lysate. It is shown that these functions require only an internal region of hsp110 that includes the predicted peptide binding domain and two immediately adjacent C-terminal domains. It is also shown that although hsp110 binds ATP, binding can be blocked by its C-terminal region.
authors.affiliation: Department of Molecular and Cellular Biophysics, Roswell Park Cancer Institute, Buffalo, New York 14263, USA.
authors.forename: H J
authors.initials: HJ
authors.lastname: Oh
authors.forename: D
authors.initials: D
authors.lastname: Easton
authors.forename: M
authors.initials: M
authors.lastname: Murawski
authors.forename: Y
authors.initials: Y
authors.lastname: Kaneko
authors.forename: J R
authors.initials: JR
authors.lastname: Subjeck
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001665:Binding Sites; D050957:HSP110 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000737:chemistry* / Q000235:genetics; D008156:Luciferases / Q000737:chemistry; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry*; D009154:Mutation; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D012156:Reticulocytes; D012995:Solubility
chemical_list: D050957:HSP110 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D018832:Molecular Chaperones; D000255:Adenosine Triphosphate; D008156:Luciferases