Glutathionylation at Cys-111 induces dissociation of wild type and FALS mutant SOD1 dimers

Rachel L. Redler, Kyle C. Wilcox, Elizabeth Anne Proctor, Lanette Fee, Michael Caplow, Nikolay Dokholyan

Research output: Contribution to journalArticle

51 Citations (Scopus)

Abstract

Mutation of the ubiquitous cytosolic enzyme Cu/Zn superoxide dismutase (SOD1) is hypothesized to cause familial amyotrophic lateral sclerosis (FALS) through structural destabilization leading to misfolding and aggregation. Considering the late onset of symptoms as well as the phenotypic variability among patients with identical SOD1 mutations, it is clear that nongenetic factor(s) impact ALS etiology and disease progression. Here we examine the effect of Cys-111 glutathionylation, a physiologically prevalent post-translational oxidative modification, on the stabilities of wild type SOD1 and two phenotypically diverse FALS mutants, A4V and I112T. Glutathionylation results in profound destabilization of SOD1 WT dimers, increasing the equilibrium dissociation constant K d to ∼10-20 μM, comparable to that of the aggressive A4V mutant. SOD1 A4V is further destabilized by glutathionylation, experiencing an ∼30-fold increase in K d. Dissociation kinetics of glutathionylated SOD1 WT and SOD1 A4V are unchanged, as measured by surface plasmon resonance, indicating that glutathionylation destabilizes these variants by decreasing association rate. In contrast, SOD1 I112T has a modestly increased dissociation rate but no change in K d when glutathionylated. Using computational structural modeling, we show that the distinct effects of glutathionylation on different SOD1 variants correspond to changes in composition of the dimer interface. Our experimental and computational results show that Cys-111 glutathionylation induces structural rearrangements that modulate stability of both wild type and FALS mutant SOD1. The distinct sensitivities of SOD1 variants to glutathionylation, a modification that acts in part as a coping mechanism for oxidative stress, suggest a novel mode by which redox regulation and aggregation propensity interact in ALS.

Original languageEnglish (US)
Pages (from-to)7057-7066
Number of pages10
JournalBiochemistry
Volume50
Issue number32
DOIs
StatePublished - Aug 16 2011

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Dimers
Agglomeration
Mutation
Oxidative stress
Surface Plasmon Resonance
Surface plasmon resonance
Post Translational Protein Processing
Oxidation-Reduction
Disease Progression
Oxidative Stress
Association reactions
Kinetics
Enzymes
Chemical analysis
Amyotrophic lateral sclerosis 1
Superoxide Dismutase
Superoxide Dismutase-1

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Redler, Rachel L. ; Wilcox, Kyle C. ; Proctor, Elizabeth Anne ; Fee, Lanette ; Caplow, Michael ; Dokholyan, Nikolay. / Glutathionylation at Cys-111 induces dissociation of wild type and FALS mutant SOD1 dimers. In: Biochemistry. 2011 ; Vol. 50, No. 32. pp. 7057-7066.
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abstract = "Mutation of the ubiquitous cytosolic enzyme Cu/Zn superoxide dismutase (SOD1) is hypothesized to cause familial amyotrophic lateral sclerosis (FALS) through structural destabilization leading to misfolding and aggregation. Considering the late onset of symptoms as well as the phenotypic variability among patients with identical SOD1 mutations, it is clear that nongenetic factor(s) impact ALS etiology and disease progression. Here we examine the effect of Cys-111 glutathionylation, a physiologically prevalent post-translational oxidative modification, on the stabilities of wild type SOD1 and two phenotypically diverse FALS mutants, A4V and I112T. Glutathionylation results in profound destabilization of SOD1 WT dimers, increasing the equilibrium dissociation constant K d to ∼10-20 μM, comparable to that of the aggressive A4V mutant. SOD1 A4V is further destabilized by glutathionylation, experiencing an ∼30-fold increase in K d. Dissociation kinetics of glutathionylated SOD1 WT and SOD1 A4V are unchanged, as measured by surface plasmon resonance, indicating that glutathionylation destabilizes these variants by decreasing association rate. In contrast, SOD1 I112T has a modestly increased dissociation rate but no change in K d when glutathionylated. Using computational structural modeling, we show that the distinct effects of glutathionylation on different SOD1 variants correspond to changes in composition of the dimer interface. Our experimental and computational results show that Cys-111 glutathionylation induces structural rearrangements that modulate stability of both wild type and FALS mutant SOD1. The distinct sensitivities of SOD1 variants to glutathionylation, a modification that acts in part as a coping mechanism for oxidative stress, suggest a novel mode by which redox regulation and aggregation propensity interact in ALS.",
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Glutathionylation at Cys-111 induces dissociation of wild type and FALS mutant SOD1 dimers. / Redler, Rachel L.; Wilcox, Kyle C.; Proctor, Elizabeth Anne; Fee, Lanette; Caplow, Michael; Dokholyan, Nikolay.

In: Biochemistry, Vol. 50, No. 32, 16.08.2011, p. 7057-7066.

Research output: Contribution to journalArticle

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T1 - Glutathionylation at Cys-111 induces dissociation of wild type and FALS mutant SOD1 dimers

AU - Redler, Rachel L.

AU - Wilcox, Kyle C.

AU - Proctor, Elizabeth Anne

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AU - Caplow, Michael

AU - Dokholyan, Nikolay

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