title: Hydrogen peroxide induces the dissociation of GroEL into monomers that can facilitate the reactivation of oxidatively inactivated rhodanese.
abstract: Although, several studies have been reported on the effects of oxidants on the structure and function of other molecular chaperones, no reports have been made so far for the chaperonin GroEL. The ability of GroEL to function under oxidative stress was investigated in this report by monitoring the effects of hydrogen peroxide (H(2)O(2)) on the structure and refolding activity of this protein. Using fluorescence spectroscopy and light scattering, we observed that GroEL showed increases in exposed hydrophobic sites and changes in tertiary and quaternary structure. Differential sedimentation, gel electrophoresis, and circular dichroism showed that H(2)O(2) treated GroEL underwent irreversible dissociation into monomers with partial loss of secondary structure. Relative to other proteins, GroEL was found to be highly resistant to oxidative damage. Interestingly, GroEL monomers produced under these conditions can facilitate the reactivation of H(2)O(2)-inactivated rhodanese but not urea-denatured rhodanese. Recovery of approximately 84% active rhodanese was obtained with either native or oxidized GroEL in the absence of GroES or ATP. In comparison, urea-denatured GroEL, BSA and the refolding mixture in the absence of proteins resulted in the recovery of 72, 50, and 49% rhodanese activity, respectively. Previous studies have shown that GroEL monomers can reactivate rhodanese. Here, we show that oxidized monomeric GroEL can reactivate oxidized rhodanese suggesting that GroEL retains the ability to protect proteins during oxidative stress.
authors.affiliation: Department of Chemistry and Biochemistry, California State University, San Marcos, CA 92096-0001, USA.
authors.forename: Girish C
authors.initials: GC
authors.lastname: Melkani
authors.forename: Case
authors.initials: C
authors.lastname: McNamara
authors.forename: Gustavo
authors.initials: G
authors.lastname: Zardeneta
authors.forename: Jose A
authors.initials: JA
authors.lastname: Mendoza
mesh_terms: D018834:Chaperonin 60 / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D004793:Enzyme Reactivators / Q000737:chemistry*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D010084:Oxidation-Reduction; D017510:Protein Folding; D017433:Protein Structure, Secondary / Q000187:drug effects*; D011994:Recombinant Proteins / Q000737:chemistry; D012542:Scattering, Radiation; D013050:Spectrometry, Fluorescence; D013499:Surface Properties / Q000187:drug effects; D013884:Thiosulfate Sulfurtransferase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D014508:Urea
chemical_list: D018834:Chaperonin 60; D004793:Enzyme Reactivators; D011994:Recombinant Proteins; D014508:Urea; D006861:Hydrogen Peroxide; D013884:Thiosulfate Sulfurtransferase