Crystal Structures of Human DJ-1 and Escherichia coli Hsp31, Which Share an Evolutionarily Conserved Domain

Lee, Sun Joo; Kim, So Jung; Kim, In Kwon; Ko, Jong Soo; Jeong, Chang-Sung; Kim, Gyung Hwa; Park, Chankyu; Kang, Shinjae; Suh, Pann Ghill; Lee, Heung Soo; Shin, Sun Mi

doi:10.1074/jbc.m304517200

Cited by 228 publications

(257 citation statements)

References 51 publications

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“…Instead DJ-1 contains the putative nucleophile Cys-106, which has the potential to form a Cys-His diad. The lack of a complete catalytic triad implies that DJ-1 may not have a protease function, a hypothesis that seemed to be supported by the failure to detect the protease activity of DJ-1 in assays using bovine serum albumin or azocasein as the substrate (27,29). On the other hand, it has been proposed that DJ-1 may have a protease function by using the Cys-His diad as the catalytic mechanism (26).…”

Section: Discussionmentioning

confidence: 92%

“…The crystal structure of DJ-1 reveals a high degree of structural similarity between DJ-1 and the bacterial protease PH1704 and E. coli chaperone protein Hsp31 (25)(26)(27)(28)(29). However, unlike PH1704 and Hsp31, DJ-1 does not have a Cys-His-(Glu/Asp) catalytic triad.…”

Section: Discussionmentioning

confidence: 99%

“…We and other groups have recently solved the crystal structure of human DJ-1, which shows that DJ-1 adopts a helix-strand-helix sandwich structure similar to the bacterial protease PH1704 and Escherichia coli chaperone protein Hsp31 (25)(26)(27)(28)(29). However, it is unclear whether DJ-1 has a protease or chaperone function because the Cys-His-(Glu/Asp) catalytic triad and the quaternary structure of PH1704 and Hsp31 are not conserved in DJ-1.…”

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confidence: 99%

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Familial Parkinson's Disease-associated L166P Mutation Disrupts DJ-1 Protein Folding and Function

Olzmann

Brown

Wilkinson

et al. 2004

Journal of Biological Chemistry

262

282

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Mutations in DJ-1, a protein of unknown function, were recently identified as the cause for an autosomal recessive, early onset form of familial Parkinson's disease. Here we report that DJ-1 is a dimeric protein that exhibits protease activity but no chaperone activity. The protease activity was abolished by mutation of Cys-106 to Ala, suggesting that DJ-1 functions as a cysteine protease. Our studies revealed that the Parkinson's disease-linked L166P mutation impaired the intrinsic folding propensity of DJ-1 protein, resulting in a spontaneously unfolded structure that was incapable of forming a homodimer with itself or a heterodimer with wild-type DJ-1. Correlating with the disruption of DJ-1 structure, the L166P mutation abolished the catalytic function of DJ-1. Furthermore, as a result of protein misfolding, the L166P mutant DJ-1 was selectively polyubiquitinated and rapidly degraded by the proteasome. Together these findings provide insights into the molecular mechanism by which loss-of-function mutations in DJ-1 lead to Parkinson's disease.

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Section: Discussionmentioning

confidence: 92%

Section: Discussionmentioning

confidence: 99%

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confidence: 99%

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Familial Parkinson's Disease-associated L166P Mutation Disrupts DJ-1 Protein Folding and Function

Olzmann

Brown

Wilkinson

et al. 2004

Journal of Biological Chemistry

262

282

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“…Crystal structures determined by several independent groups (21)(22)(23)(24)(25) show that C106 is located at a solventexposed sharp bend between a -strand and an R-helix called the "nucleophile elbow" (26). The backbone φ and ψ angles for the nucleophile elbow cysteine fall in the unfavorable region of Ramachandran space for every structurally characterized member of the DJ-1 superfamily (27), possibly contributing to its reactivity.…”

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confidence: 99%

Cysteine pK_a Depression by a Protonated Glutamic Acid in Human DJ-1

et al. 2008

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Human DJ-1, a disease-associated protein that protects cells from oxidative stress, contains an oxidation-sensitive cysteine (C106) that is essential for its cytoprotective activity. The origin of C106 reactivity is obscure, due in part to the absence of an experimentally determined pK a value for this residue. We have used atomic-resolution X-ray crystallography and UV spectroscopy to show that C106 has a depressed pK a of 5.4 ( 0.1 and that the C106 thiolate accepts a hydrogen bond from a protonated glutamic acid side chain (E18). X-ray crystal structures and cysteine pK a analysis of several site-directed substitutions at residue 18 demonstrate that the protonated carboxylic acid side chain of E18 is required for the maximal stabilization of the C106 thiolate. A nearby arginine residue (R48) participates in a guanidinium stacking interaction with R28 from the other monomer in the DJ-1 dimer and elevates the pK a of C106 by binding an anion that electrostatically suppresses thiol ionization. Our results show that the ionizable residues (E18, R48, and R28) surrounding C106 affect its pK a in a way that is contrary to expectations based on the typical ionization behavior of glutamic acid and arginine. Lastly, a search of the Protein Data Bank (PDB) produces several candidate hydrogen-bonded aspartic/glutamic acid-cysteine interactions, which we propose are particularly common in the DJ-1 superfamily.Cysteine thiolates are potent nucleophiles that are used by many proteins for catalysis, metal binding, or to facilitate post-translational modification. However, the solution pK a value of cysteine (8.3) is not within the optimal physiological pH range of most proteins. To render cysteine residues reactive, the thiol pK a must be depressed by stabilization of the conjugate thiolate anion by the protein environment. Multiple types of stabilizing interactions can decrease cysteine pK a values, including electrostatic complementarity with nearby cations (1, 2), the R-helix macropole (3), and hydrogen bonding to the thiol (4).The Parkinsonism-associated protein DJ-1 contains an oxidation-sensitive cysteine residue (C106 in human DJ-1) of unknown pK a that is required for the cytoprotective activity of the protein (5, 6). DJ-1 is a homodimeric protein of 189 amino acid monomers that is found both in the cytoplasm (7) and in the mitochondria (5, 8) and protects cells from various types of oxidative stress (5,(9)(10)(11)(12)(13)(14)(15)(16)(17)(18). Although C106 has been shown to be essential for the protective function of DJ-1 in cell culture (5, 16) and Drosophila melanogaster (6), the specific biochemical activity of the protein that requires this cysteine residue is uncertain. Identifying the structural determinants of C106 ionization (and hence reactivity) is an essential first step in understanding how this conserved cysteine residue contributes to the neuroprotective activity of DJ-1.The reactive cysteine in human DJ-1 is one of the most highly conserved residues in the DJ-1 superfamily (19,20). Crystal s...

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“…It is highly conserved from human to Escherichia coli and is localized in the cytoplasm, nucleus, and mitochondria (Bonifati et al, 2003;Gupta et al, 2008;Lee et al, 2003;Wilson et al, 2004). This protein has multiple functions including antioxidant activity, chaperonelike properties, and transcriptional regulation (Menzies et al, 2005;Xu et al, 2005;Zhou et al, 2006).…”

Section: Introductionmentioning

confidence: 99%

Transduced Tat-DJ-1 Protein Protects against Oxidative Stress-Induced SH-SY5Y Cell Death and Parkinson Disease in a Mouse Model

Jeong

Kim

Woo

et al. 2012

Molecules and Cells

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Parkinson's disease (PD) is a well known neurodegenerative disorder characterized by selective loss of dopaminergic neurons in the substantia nigra pars compact (SN). Although the exact mechanism remains unclear, oxidative stress plays a critical role in the pathogenesis of PD. DJ-1 is a multifunctional protein, a potent antioxidant and chaperone, the loss of function of which is linked to the autosomal recessive early onset of PD. Therefore, we investigated the protective effects of DJ-1 protein against SH-SY5Y cells and in a PD mouse model using a cell permeable Tat-DJ-1 protein. Tat-DJ-1 protein rapidly transduced into the cells and showed a protective effect on 6-hydroxydopamine (6-OHDA)-induced neuronal cell death by reducing the reactive oxygen species (ROS). In addition, we found that Tat-DJ-1 protein protects against dopaminergic neuronal cell death in 1-methyl-4-phenyl-1,2,3,6,-tetrahydropyridine (MPTP)-induced PD mouse models. These results suggest that Tat-DJ-1 protein provides a potential therapeutic strategy for against ROS related human diseases including PD.

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Crystal Structures of Human DJ-1 and Escherichia coli Hsp31, Which Share an Evolutionarily Conserved Domain

Cited by 228 publications

References 51 publications

Familial Parkinson's Disease-associated L166P Mutation Disrupts DJ-1 Protein Folding and Function

Familial Parkinson's Disease-associated L166P Mutation Disrupts DJ-1 Protein Folding and Function

Cysteine pK_a Depression by a Protonated Glutamic Acid in Human DJ-1

Transduced Tat-DJ-1 Protein Protects against Oxidative Stress-Induced SH-SY5Y Cell Death and Parkinson Disease in a Mouse Model

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Crystal Structures of Human DJ-1 and Escherichia coli Hsp31, Which Share an Evolutionarily Conserved Domain

Cited by 228 publications

References 51 publications

Familial Parkinson's Disease-associated L166P Mutation Disrupts DJ-1 Protein Folding and Function

Familial Parkinson's Disease-associated L166P Mutation Disrupts DJ-1 Protein Folding and Function

Cysteine pKa Depression by a Protonated Glutamic Acid in Human DJ-1

Transduced Tat-DJ-1 Protein Protects against Oxidative Stress-Induced SH-SY5Y Cell Death and Parkinson Disease in a Mouse Model

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Cysteine pK_a Depression by a Protonated Glutamic Acid in Human DJ-1