title: Unfolding and disassembly of the chaperonin GroEL occurs via a tetradecameric intermediate with a folded equatorial domain.
abstract: The chaperonin GroEL is a homotetradecamer in which the subunits (M(r) 57 000) are joined through noncovalent forces. This study reports on the unfolding and disassembly of GroEL in guanidine hydrochloride and urea. Kinetic and equilibrium measurements were made using amide hydrogen exchange/mass spectrometry, light scattering, and size-exclusion chromatography. Hydrogen exchange in GroEL destabilized in 1.8 M GdHCl (the unfolding midpoint is 1.2 M GdHCl) shows that the apical and intermediate domains unfold 3.1 times faster than the equatorial domain. Light scattering measurements made under the same conditions show that disassembly of the native GroEL tetradecamer occurs at the same rate as unfolding of the equatorial domain. This study of the kinetics of GroEL unfolding and disassembly demonstrates the existence of an intermediate that was identified as a tetradecamer with the apical and intermediate domains unfolded. Although this intermediate was easily detected in dynamic unfolding measurements, its population in equilibrium measurements at the midpoint for GroEL unfolding was too small to be detected. This study of GroEL unfolding and disassembly points to features that may be important in the folding and assembly of the GroEL macroassembly.
authors.affiliation: Department of Chemistry and Eppley Institute for Research in Cancer and Allied Diseases, University of Nebraska, Lincoln, Nebraska 68588-0304, USA.
authors.forename: J
authors.initials: J
authors.lastname: Chen
authors.forename: D L
authors.initials: DL
authors.lastname: Smith
mesh_terms: D000595:Amino Acid Sequence; D018834:Chaperonin 60 / Q000737:chemistry* / Q000378:metabolism*; D002850:Chromatography, Gel; D003903:Deuterium / Q000378:metabolism; D017666:Deuterium Oxide / Q000378:metabolism; D004926:Escherichia coli; D019791:Guanidine / Q000494:pharmacology; D007700:Kinetics; D008027:Light; D013058:Mass Spectrometry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010434:Pepsin A / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism; D011489:Protein Denaturation / Q000187:drug effects; D017510:Protein Folding; D020836:Protein Structure, Quaternary / Q000187:drug effects; D017434:Protein Structure, Tertiary / Q000187:drug effects; D012542:Scattering, Radiation; D013816:Thermodynamics; D014508:Urea / Q000494:pharmacology
chemical_list: D018834:Chaperonin 60; D010446:Peptide Fragments; D014508:Urea; D003903:Deuterium; D010434:Pepsin A; D017666:Deuterium Oxide; D019791:Guanidine