title: Denaturation by urea and renaturation of 20 beta-hydroxysteroid dehydrogenase studied by high-performance size exclusion chromatography.
abstract: The denaturation by urea and renaturation of 20 beta-hydroxysteroid dehydrogenase, a tetrameric enzyme consisting of four identical subunits, were followed by high-performance size exclusion chromatography to detect intermediates in the processes. During the denaturation process no intermediate form (structured monomers or dimers) between the tetramer and the denatured monomer was observed. During the renaturation process, carried out either with or without NADH, high molecular weight aggregates, native tetramers, and low molecular weight intermediates were evidenced and quantified. The contemporaneous measurement of recovery of activity unambiguously demonstrated that the tetrameric structure is essential for enzymatic activity.
authors.affiliation: Istituto di Chimica degli Ormoni del C.N.R., Milan, Italy.
authors.forename: G
authors.initials: G
authors.lastname: Carrea
authors.forename: R
authors.initials: R
authors.lastname: Longhi
authors.forename: G
authors.initials: G
authors.lastname: Mazzola
authors.forename: P
authors.initials: P
authors.lastname: Pasta
authors.forename: G
authors.initials: G
authors.lastname: Vecchio
mesh_terms: D015089:20-Hydroxysteroid Dehydrogenases / Q000032:analysis*; D002850:Chromatography, Gel / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003349:Cortisone Reductase / Q000032:analysis*; D011489:Protein Denaturation / Q000187:drug effects; D013379:Substrate Specificity / Q000187:drug effects; D014508:Urea / Q000494:pharmacology*
chemical_list: D014508:Urea; D015089:20-Hydroxysteroid Dehydrogenases; D003349:Cortisone Reductase