TY - JOUR
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 A.
AU - Fee, Lanette
AU - Caplow, Michael
AU - Dokholyan, Nikolay V.
PY - 2011/8/16
Y1 - 2011/8/16
N2 - 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.
AB - 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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U2 - 10.1021/bi200614y
DO - 10.1021/bi200614y
M3 - Article
C2 - 21739997
AN - SCOPUS:80051503291
SN - 0006-2960
VL - 50
SP - 7057
EP - 7066
JO - Biochemistry
JF - Biochemistry
IS - 32
ER -