Protection of GroEL by its methionine residues against oxidation by hydrogen peroxide.
GroEL undergoes an important functional and structural transition when oxidized with hydrogen peroxide (H2O2) concentrations between 15 and 20mM. When GroEL was incubated for 3h with 15 mM H2O2, it retained its quaternary structure, chaperone and ATPase activities. Under these conditions, GroEL's cysteine and tyrosine residues remained intact. However, all the methionine residues of the molecular chaperone were oxidized to the corresponding methionine-sulfoxides under these conditions. The oxidation of the methionine residues was verified by the inability of cyanogen bromide to cleave at the carboxyl side of the modified methionine residues. The role for the proportionately large number (23) of methionine residues in GroEL has not been identified. Methionine residues have been reported to have an antioxidant activity in proteins against a variety of oxidants produced in biological systems including H2O2. The carboxyl-terminal domain of GroEL is rich in methionine residues and we hypothesized that these residues are involved in the protection of GroEL's functional structure by scavenging H2O2. When GroEL was further incubated for the same time, but with increasing concentrations of H2O2 (>15 mM), the oxidation of GroEL's cysteine residues and a significant decrease of the tyrosine fluorescence due to the formation of dityrosines were observed. Also, at these higher concentrations of H2O2, the inability of GroEL to hydrolyze ATP and to assist the refolding of urea-unfolded rhodanese was observed.
Melkani
Girish C
GC
Department of Chemistry and Biochemistry, California State University at San Marcos, San Marcos, CA 92096-0001, USA.
Kestetter
Justin
J
Sielaff
Robin
R
Zardeneta
Gustavo
G
Mendoza
Jose A
JA
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D018834:Chaperonin 60 / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D003545:Cysteine / Q000737:chemistry / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D006868:Hydrolysis / Q000187:drug effects; D008715:Methionine / Q000737:chemistry* / Q000378:metabolism; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D016877:Oxidants / Q000494:pharmacology; D010084:Oxidation-Reduction / Q000187:drug effects; D017510:Protein Folding; D020836:Protein Structure, Quaternary / Q000187:drug effects; D017433:Protein Structure, Secondary / Q000187:drug effects; D013884:Thiosulfate Sulfurtransferase / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry / Q000378:metabolism
D018834:Chaperonin 60; D018832:Molecular Chaperones; D016877:Oxidants; D014443:Tyrosine; D000255:Adenosine Triphosphate; D008715:Methionine; D006861:Hydrogen Peroxide; D013884:Thiosulfate Sulfurtransferase; D000251:Adenosine Triphosphatases; D003545:Cysteine