title: Dissociation and unfolding of the pyruvate dehydrogenase complex by guanidinium chloride.
abstract: The effect of guanidinium chloride (GdnHCl) on the pyruvate dehydrogenase complex (PDC) from bovine heart and its constituent enzymes has been studied. The overall activity of the complex is lost reversibly at low levels of GdnHCl (0.2 M) which cause 40-50% inactivation but no loss of overall secondary or tertiary structures of the individual enzymes; the inactivation of the complex is shown to be caused by dissociation of the E1 and E3 components from the E2/X core assembly. This provides an improved procedure for controlled dissociation of the complex and efficient recovery of its component enzymes in their native states. Higher concentrations of GdnHCl (up to 4 M) lead to the unfolding and irreversible inactivation of the separate enzymes of the complex with the E2/X core proving the most resistant to GdnHCl-induced unfolding. Neither the 60-meric E2/X core assembly nor the dimeric E3 component are dissociated into monomers in the presence of 6 M GdnHCl; the latter enzyme forms higher-M(r) aggregates under these conditions.
authors.affiliation: Division of Biochemistry and Molecular Biology, University of Glasgow, Scotland, U.K.
authors.forename: S M
authors.initials: SM
authors.lastname: West
authors.forename: J E
authors.initials: JE
authors.lastname: Rice
authors.forename: E S
authors.initials: ES
authors.lastname: Beaumont
authors.forename: S M
authors.initials: SM
authors.lastname: Kelly
authors.forename: N C
authors.initials: NC
authors.lastname: Price
authors.forename: J G
authors.initials: JG
authors.lastname: Lindsay
mesh_terms: D000818:Animals; D002417:Cattle; D002850:Chromatography, Gel; D002942:Circular Dichroism; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005453:Fluorescence; D019791:Guanidine; D006146:Guanidines / Q000494:pharmacology*; D009206:Myocardium / Q000201:enzymology; D017510:Protein Folding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011768:Pyruvate Dehydrogenase Complex / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism
chemical_list: D006146:Guanidines; D011768:Pyruvate Dehydrogenase Complex; D019791:Guanidine