Effect of single amino acid substitution on oxidative modifications of the Parkinson's disease-related protein, DJ-1.

Ashraf G. Madian, Jagadish Hindupur, John D. Hulleman, Naomi Diaz-Maldonado, Vartika R. Mishra, Emmanuel Guigard, Cyril M. Kay, Jean Christophe Rochet, Fred E. Regnier

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Abstract

Mutations in the gene encoding DJ-1 have been identified in patients with familial Parkinson's disease (PD) and are thought to inactivate a neuroprotective function. Oxidation of the sulfhydryl group to a sulfinic acid on cysteine residue C106 of DJ-1 yields the "2O " form, a variant of the protein with enhanced neuroprotective function. We hypothesized that some familial mutations disrupt DJ-1 activity by interfering with conversion of the protein to the 2O form. To address this hypothesis, we developed a novel quantitative mass spectrometry approach to measure relative changes in oxidation at specific sites in mutant DJ-1 as compared with the wild-type protein. Treatment of recombinant wild-type DJ-1 with a 10-fold molar excess of H(2)O(2) resulted in a robust oxidation of C106 to the sulfinic acid, whereas this modification was not detected in a sample of the familial PD mutant M26I exposed to identical conditions. Methionine oxidized isoforms of wild-type DJ-1 were depleted, presumably as a result of misfolding and aggregation, under conditions that normally promote conversion of the protein to the 2O form. These data suggest that the M26I familial substitution and methionine oxidation characteristic of sporadic PD may disrupt DJ-1 function by disfavoring a site-specific modification required for optimal neuroprotective activity. Our findings indicate that a single amino acid substitution can markedly alter a protein's ability to undergo oxidative modification, and they imply that stimulating the conversion of DJ-1 to the 2O form may be therapeutically beneficial in familial or sporadic PD.

Original languageEnglish (US)
JournalMolecular & cellular proteomics : MCP
Volume11
Issue number2
DOIs
StatePublished - Feb 2012

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Amino Acid Substitution
Parkinson Disease
Substitution reactions
Amino Acids
Sulfinic Acids
Oxidation
Proteins
Methionine
Mutation
Gene encoding
Mass spectrometry
Cysteine
Mass Spectrometry
Protein Isoforms
Agglomeration
Protein Deglycase DJ-1
Genes

ASJC Scopus subject areas

  • Medicine(all)

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Effect of single amino acid substitution on oxidative modifications of the Parkinson's disease-related protein, DJ-1. / Madian, Ashraf G.; Hindupur, Jagadish; Hulleman, John D.; Diaz-Maldonado, Naomi; Mishra, Vartika R.; Guigard, Emmanuel; Kay, Cyril M.; Rochet, Jean Christophe; Regnier, Fred E.

In: Molecular & cellular proteomics : MCP, Vol. 11, No. 2, 02.2012.

Research output: Contribution to journalArticle

Madian, Ashraf G. ; Hindupur, Jagadish ; Hulleman, John D. ; Diaz-Maldonado, Naomi ; Mishra, Vartika R. ; Guigard, Emmanuel ; Kay, Cyril M. ; Rochet, Jean Christophe ; Regnier, Fred E. / Effect of single amino acid substitution on oxidative modifications of the Parkinson's disease-related protein, DJ-1. In: Molecular & cellular proteomics : MCP. 2012 ; Vol. 11, No. 2.
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