Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells

Pokrishevsky, Edward; McAlary, Luke; Farrawell, Natalie E.; Zhao, Beibei; Sher, Mine; Yerbury, Justin J.; Cashman, Neil R.

doi:10.1038/s41598-018-32835-y

Cited by 40 publications

(90 citation statements)

References 59 publications

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“…Indeed, only a few simian species and the African elephant are known to incorporate tryptophan at this position while a conservative phenylalanine substitution is found at the equivalent position in human extracellular CuZnSOD (SOD3) indicating hydrophobicity at this site has some functionality. Accordingly, Pokrishevsky et al (2018) showed that substitution of Trp32 for the more frequently found serine decreases mutant and wild-type SOD1 stability and resistance to proteolytic digestion.…”

Section: Small Moleculesmentioning

confidence: 99%

The biophysics of superoxide dismutase-1 and amyotrophic lateral sclerosis

Wright

Antonyuk

Hasnain

2019

Quart. Rev. Biophys.

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Few proteins have come under such intense scrutiny as superoxide dismutase-1 (SOD1). For almost a century, scientists have dissected its form, function and then later its malfunction in the neurodegenerative disease amyotrophic lateral sclerosis (ALS). We now know SOD1 is a zinc and copper metalloenzyme that clears superoxide as part of our antioxidant defence and respiratory regulation systems. The possibility of reduced structural integrity was suggested by the first crystal structures of human SOD1 even before deleterious mutations in thesod1gene were linked to the ALS. This concept evolved in the intervening years as an impressive array of biophysical studies examined the characteristics of mutant SOD1 in great detail. We now recognise how ALS-related mutations perturb the SOD1 maturation processes, reduce its ability to fold and reduce its thermal stability and half-life. Mutant SOD1 is therefore predisposed to monomerisation, non-canonical self-interactions, the formation of small misfolded oligomers and ultimately accumulation in the tell-tale insoluble inclusions found within the neurons of ALS patients. We have also seen that several post-translational modifications could push wild-type SOD1 down this toxic pathway. Recently we have come to view ALS as a prion-like disease where both the symptoms, and indeed SOD1 misfolding itself, are transmitted to neighbouring cells. This raises the possibility of intervention after the initial disease presentation. Several small-molecule and biologic-based strategies have been devised which directly target the SOD1 molecule to change the behaviour thought to be responsible for ALS. Here we provide a comprehensive review of the many biophysical advances that sculpted our view of SOD1 biology and the recent work that aims to apply this knowledge for therapeutic outcomes in ALS.

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Section: Small Moleculesmentioning

confidence: 99%

The biophysics of superoxide dismutase-1 and amyotrophic lateral sclerosis

Wright

Antonyuk

Hasnain

2019

Quart. Rev. Biophys.

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“…Modulation of ALS mutant SOD1 aggregation by substitution of Cys 111 to Ser has also been described [70][71][72]. The mutation of Trp 32 to Ser potently inhibited the ability of misfolded WT SOD1 to propagate between cells in culture [57,68,69]. Small molecules that bind near Trp 32 can block the seeding of mutant SOD1 aggregation in cell culture models [68,69,73].…”

Section: Introductionmentioning

confidence: 96%

“…Prior studies of ALS mutant SOD1 aggregation have shown that mutation of the unique Trp residue at position 32 of SOD1 to Ser or Phe can act in cis to suppress aggregation [38,[67][68][69]. Modulation of ALS mutant SOD1 aggregation by substitution of Cys 111 to Ser has also been described [70][71][72].…”

Section: Introductionmentioning

confidence: 98%

“…The mutation of Trp 32 to Ser potently inhibited the ability of misfolded WT SOD1 to propagate between cells in culture [57,68,69]. Small molecules that bind near Trp 32 can block the seeding of mutant SOD1 aggregation in cell culture models [68,69,73]. In the present study, we used a combination of cell culture models and in vivo seeding models to examine the role of Trp 32 in modulating mutant SOD1 aggregation.…”

Section: Introductionmentioning

confidence: 99%

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Tryptophan residue 32 in human Cu-Zn superoxide dismutase modulates prion-like propagation and strain selection

et al. 2020

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Mutations in Cu/Zn superoxide dismutase 1 (SOD1) associated with familial amyotrophic lateral sclerosis cause the protein to aggregate via a prion-like process in which soluble molecules are recruited to aggregates by conformational templating. These misfolded SOD1 proteins can propagate aggregation-inducing conformations across cellular membranes. Prior studies demonstrated that mutation of a Trp (W) residue at position 32 to Ser (S) suppresses the propagation of misfolded conformations between cells, whereas other studies have shown that mutation of Trp 32 to Phe (F), or Cys 111 to Ser, can act in cis to attenuate aggregation of mutant SOD1. By expressing mutant SOD1 fused with yellow fluorescent protein (YFP), we compared the relative ability of these mutations to modulate the formation of inclusions by ALS-mutant SOD1 (G93A and G85R). Only mutation of Trp 32 to Ser persistently reduced the formation of the amorphous inclusions that form in these cells, consistent with the idea that a Ser at position 32 inhibits templated propagation of aggregation prone conformations. To further test this idea, we produced aggregated fibrils of recombinant SOD1-W32S in vitro and injected them into the spinal cords of newborn mice expressing G85R-SOD1: YFP. The injected mice developed an earlier onset paralysis with a frequency similar to mice injected with WT SOD1 fibrils, generating a strain of misfolded SOD1 that produced highly fibrillar inclusion pathology. These findings suggest that the effect of Trp 32 in modulating the propagation of misfolded SOD1 conformations may be dependent upon the "strain" of the conformer that is propagating.

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“…We have found that the prion-like seeding and propagation of human wild-type SOD1 misfolding requires a tryptophan-tryptophan interaction in neighboring SOD1 molecules mediated by the single tryptophan at position 32 in SOD1 (Trp32) (10)(11)(12). Also, Trp32 mediates SOD1 aggregation in cultured cells, as well as zebrafish axonopathy in vivo (13,14). Furthermore, drugs that interact with Trp32 (15,16) can block propagated SOD1 aggregation in cell cultures and its toxicity in vivo (13,14).…”

Section: Introductionmentioning

confidence: 99%

Tryptophan residues in TDP-43 and SOD1 mediate the cross-seeding and toxicity of SOD1

Pokrishevsky

DuVal

McAlary

et al. 2020

Preprint

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Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease of motor neurons. Neuronal superoxide dismutase-1 (SOD1) inclusion bodies are characteristic of familial ALS with SOD1 mutations, while a hallmark of sporadic ALS is inclusions containing aggregated wild-type TAR DNA-binding protein 43 (TDP-43). Co-expression of mutant or wild-type TDP-43 with SOD1 leads to misfolding of endogenous SOD1 and aggregation of SOD1 reporter protein G85R-GFP in HEK293FT cells, and promotes synergistic axonopathy in zebrafish. This pathological interaction is dependent upon natively solvent-exposed tryptophans in SOD1 (tryptophan-32) and TDP-43 RRM1 (tryptophan-172), in concert with natively sequestered TDP-43 N-terminal domain tryptophan-68. TDP-43 RRM1 intrabodies reduce wild-type SOD1 misfolding in HEK293FT cells, via blocking tryptophan-172. Tryptophan-68 becomes antibody-accessible in aggregated TDP-43 in sporadic ALS motor neurons and cell culture. 5-fluorouridine inhibits TDP-43-induced G85R-GFP SOD1 aggregation in HEK293FT cells, and ameliorates axonopathy in zebrafish, via its interaction with SOD1 tryptophan-32. Collectively, our results establish a novel and potentially druggable tryptophan-mediated mechanism whereby two principal ALS disease effector proteins might directly interact in disease.

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Tryptophan 32-mediated SOD1 aggregation is attenuated by pyrimidine-like compounds in living cells

Cited by 40 publications

References 59 publications

The biophysics of superoxide dismutase-1 and amyotrophic lateral sclerosis

The biophysics of superoxide dismutase-1 and amyotrophic lateral sclerosis

Tryptophan residue 32 in human Cu-Zn superoxide dismutase modulates prion-like propagation and strain selection

Tryptophan residues in TDP-43 and SOD1 mediate the cross-seeding and toxicity of SOD1

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