15N CEST data and traditional model-free analysis capture fast internal dynamics of DJ-1

Catazaro, Jonathan; Andrews, Tessa; Milkovic, Nicole M.; Lin, Jiusheng; Lowe, Austin J.; Wilson, Mark A.; Powers, Robert

doi:10.1016/j.ab.2017.11.012

Cited by 6 publications

(10 citation statements)

References 18 publications

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“…Two-dimensional (2D) 1 H- 15 N heteronuclear single quantum coherence (HSQC) spectra were collected at 37 °C as previously described 42,43 from uniformly 15 N labeled DJ-1 that was either reduced or overoxidized to Cys106-SO 3 − The 2D 1 H- 15 N HSQC spectra were collected on a 700 MHz Bruker Avance III spectrometer equipped with a 5 mm quadruple resonance QCI-P cryoprobe ( 1 H, 13 C, 15 N, and 31 P) with z -axis gradients. Order parameters ( S 2 ) were calculated from chemical exchange saturation transfer (CEST)-derived R 1 and R 2 relaxation rates and heteronuclear NOE values using FAST-Modelfree 44 as previously described 42 .…”

Section: Methodsmentioning

confidence: 99%

The effect of cysteine oxidation on DJ-1 cytoprotective function in human alveolar type II cells

et al. 2019

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DJ-1 is a multifunctional protein with cytoprotective functions. It is localized in the cytoplasm, nucleus, and mitochondria. The conserved cysteine residue at position 106 (Cys106) within DJ-1 serves as a sensor of redox state and can be oxidized to both the sulfinate (-SO 2 − ) and sulfonate (-SO 3 − ) forms. DJ-1 with Cys106-SO 2 − has cytoprotective activity but high levels of reactive oxygen species can induce its overoxidation to Cys106-SO 3 − . We found increased oxidative stress in alveolar type II (ATII) cells isolated from emphysema patients as determined by 4-HNE expression. DJ-1 with Cys106-SO 3 − was detected in these cells by mass spectrometry analysis. Moreover, ubiquitination of Cys106-SO 3 − DJ-1 was identified, which suggests that this oxidized isoform is targeted for proteasomal destruction. Furthermore, we performed controlled oxidation using H 2 O 2 in A549 cells with DJ-1 knockout generated using CRISPR-Cas9 strategy. Lack of DJ-1 sensitized cells to apoptosis induced by H 2 O 2 as detected using Annexin V and propidium iodide by flow cytometry analysis. This treatment also decreased both mitochondrial DNA amount and mitochondrial ND1 (NADH dehydrogenase 1, subunit 1) gene expression, as well as increased mitochondrial DNA damage. Consistent with the decreased cytoprotective function of overoxidized DJ-1, recombinant Cys106-SO 3 − DJ-1 exhibited a loss of its thermal unfolding transition, mild diminution of secondary structure in CD spectroscopy, and an increase in picosecond–nanosecond timescale dynamics as determined using NMR. Altogether, our data indicate that very high oxidative stress in ATII cells in emphysema patients induces DJ-1 overoxidation to the Cys106-SO 3 − form, leading to increased protein flexibility and loss of its cytoprotective function, which may contribute to this disease pathogenesis.

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Section: Methodsmentioning

confidence: 99%

The effect of cysteine oxidation on DJ-1 cytoprotective function in human alveolar type II cells

et al. 2019

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“…The reduced form of DJ-1 (DJ-1 Cys106-SH) can be oxidized to a sulfinic acid form (DJ-1 Cys106-SO 2 H) and a sulfonic acid form (DJ-1 Cys106-SO 3 H) in the presence of moderate or high oxidative stress paradigms (Figure 1). While the reduced and sulfinic DJ-1 forms are stable, the sulfonic form of DJ-1 is unstable and prone to aggregate formation [47,48,49].…”

Section: Introductionmentioning

confidence: 99%

DJ-1 in Parkinson’s Disease: Clinical Insights and Therapeutic Perspectives

Repici

Giorgini

2019

JCM

131

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Mutations in the protein DJ-1 cause autosomal recessive forms of Parkinson’s disease (PD) and oxidized DJ-1 is found in the brains of idiopathic PD individuals. While several functions have been ascribed to DJ-1 (most notably protection from oxidative stress), its contribution to PD pathogenesis is not yet clear. Here we provide an overview of the clinical research to date on DJ-1 and the current state of knowledge regarding DJ-1 characterization in the human brain. The relevance of DJ-1 as a PD biomarker is also discussed, as are studies exploring DJ-1 as a possible therapeutic target for PD and neurodegeneration.

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“…Oxidation also regulates DJ-1 subcellular distribution so that it can access binding partners localized to organelles, including stress granules and mitochondria (reviewed). However, further C106 oxidation to sulfonic acid form (i.e., C106-SO 3 H) owing to chronic oxidative stress destabilizes dimeric DJ-1 structure and leads to irreversible inactivation. , Here, we found that oxidation incompetent C106A mutant retains full tau chaperone activity, indicating that oxidation is not required for interaction with this specific client. These data predict that DJ-1 can protect full-length tau protein against aggregation under basal conditions in the absence of oxidative stress.…”

Section: Discussionmentioning

confidence: 99%

DJ-1 Molecular Chaperone Activity Depresses Tau Aggregation Propensity through Interaction with Monomers

et al. 2023

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Tau aggregate-bearing lesions are pathological markers and potential mediators of tauopathic neurodegenerative diseases, including Alzheimer’s disease. The molecular chaperone DJ-1 colocalizes with tau pathology in these disorders, but it has been unclear what functional link exists between them. In this study, we examined the consequences of tau/DJ-1 interaction as isolated proteins in vitro. When added to full-length 2N4R tau under aggregation-promoting conditions, DJ-1 inhibited both the rate and extent of filament formation in a concentration-dependent manner. Inhibitory activity was low affinity, did not require ATP, and was not affected by substituting oxidation incompetent missense mutation C106A for wild-type DJ-1. In contrast, missense mutations previously linked to familial Parkinson’s disease and loss of α-synuclein chaperone activity, M26I and E64D, displayed diminished tau chaperone activity relative to wild-type DJ-1. Although DJ-1 directly bound the isolated microtubule-binding repeat region of tau protein, exposure of preformed tau seeds to DJ-1 did not diminish seeding activity in a biosensor cell model. These data reveal DJ-1 to be a holdase chaperone capable of engaging tau as a client in addition to α-synuclein. Our findings support a role for DJ-1 as part of an endogenous defense against the aggregation of these intrinsically disordered proteins.

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15N CEST data and traditional model-free analysis capture fast internal dynamics of DJ-1

Cited by 6 publications

References 18 publications

The effect of cysteine oxidation on DJ-1 cytoprotective function in human alveolar type II cells

The effect of cysteine oxidation on DJ-1 cytoprotective function in human alveolar type II cells

DJ-1 in Parkinson’s Disease: Clinical Insights and Therapeutic Perspectives

DJ-1 Molecular Chaperone Activity Depresses Tau Aggregation Propensity through Interaction with Monomers

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