title: Development and optimization of a useful assay for determining Hsp90's inherent ATPase activity.
abstract: The Hsp90 molecular chaperone is responsible for the conformational maturation of nascent polypeptides and the rematuration of denatured proteins. Inhibition of Hsp90 represents a promising approach towards the treatment of cancer because numerous signaling cascades can be simultaneously targeted by disruption of the Hsp90-mediated process. Hsp90's ATPase activity is essential to the Hsp90-mediated protein folding process, consequently, a coupled assay was developed and optimized for determination of Hsp90's inherent ATPase activity. Using maltose phosphorylase, glucose oxidase, and horseradish peroxidase as components of this assay, a highly reproducible assay with a Z-factor of 0.87 has been produced.
authors.affiliation: The Department of Medicinal Chemistry, The University of Kansas, Malott 4070, Lawrence, 66045-7563, USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Avila
authors.forename: Boris A
authors.initials: BA
authors.lastname: Kornilayev
authors.forename: Brian S J
authors.initials: BS
authors.lastname: Blagg
mesh_terms: D000251:Adenosine Triphosphatases / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism* / Q000494:pharmacology; D002384:Catalysis; D005949:Glucose Oxidase / Q000378:metabolism; D005964:Glucosyltransferases / Q000378:metabolism; D018841:HSP90 Heat-Shock Proteins / Q000032:analysis / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D006735:Horseradish Peroxidase / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology; D015394:Molecular Structure; D015203:Reproducibility of Results; D013696:Temperature
chemical_list: D018841:HSP90 Heat-Shock Proteins; D000255:Adenosine Triphosphate; D006861:Hydrogen Peroxide; D005949:Glucose Oxidase; D006735:Horseradish Peroxidase; D005964:Glucosyltransferases; C021535:maltose phosphorylase; D000251:Adenosine Triphosphatases