Identification of Protein Interfaces between α-Synuclein, the Principal Component of Lewy Bodies in Parkinson Disease, and the Molecular Chaperones Human Hsc70 and the Yeast Ssa1p

Redeker, Virginie; Pemberton, Samantha; Bienvenut, Willy V.; Bousset, Luc; Melki, Ronald

doi:10.1074/jbc.m112.387530

Cited by 45 publications

(61 citation statements)

References 42 publications

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“…Scale bar, 20 m. bitrap and both MS and MS/MS data analysis, as described under "Experimental Procedures" and in Ref. 29. Seven Hsc70-HttEx1Q25 cross-links were identified (Table 2).…”

Section: Hsc70 Effects On Httex1qn Assembly Are Independent Ofmentioning

confidence: 99%

“…We next mapped the surface interfaces between Hsc70 and HttEx1Qn using chemical cross-linking with the homobifunctional NHS-ester BS3 cross-linker and mass spectrometry, using a strategy that we developed previously to assess Hsc70 interaction with another client protein and described in detail previously (29). Hsc70 interacts in a similar way with all of the soluble forms of HttEx1Qn we tested (Fig.…”

Section: Hsc70 Effects On Httex1qn Assembly Are Independent Ofmentioning

confidence: 99%

“…Tryptic peptides were analyzed by nanoLC-LTQ-Orbitrap mass spectrometry analysis, and the nanoLC-MS/MS data were processed as described previously (29), except that the data analysis included lysine, serine, threonine, tyrosine, and the N-terminal amino acid residues as possible cross-linked sites (30) and that the cross-linker was BS3-d0 and BS3-d4. Briefly, nanoLC-MS data were deisotoped using the Decon2LS software (available at the Pacific Northwest National Laboratory Web site).…”

Section: Expression and Purification Of Recombinant Polypeptides And mentioning

confidence: 99%

“…Peptide Preparation and NanoLC-Linear Ion Trap (LTQ)-Orbitrap Mass Spectrometry Analysis-Cross-linked protein complex spots separated by two-dimensional gel electrophoresis were excised and subjected to tryptic or GluC digestion using the Progest robot, and the digested peptides were extracted as described previously (29). Tryptic peptides were analyzed by nanoLC-LTQ-Orbitrap mass spectrometry analysis, and the nanoLC-MS/MS data were processed as described previously (29), except that the data analysis included lysine, serine, threonine, tyrosine, and the N-terminal amino acid residues as possible cross-linked sites (30) and that the cross-linker was BS3-d0 and BS3-d4.…”

Section: Expression and Purification Of Recombinant Polypeptides And mentioning

confidence: 99%

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Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Monsellier

Redeker

Ruiz-Arlandis

et al. 2015

Journal of Biological Chemistry

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106

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Background: Hsc70 has an alleviating effect on the toxicity of polyglutamine (polyQ)-containing proteins in vivo. Results: Hsc70 binds specifically the N-terminal flank of huntingtin exon 1. Conclusion: Hsc70 interaction with huntingtin exon 1 N-terminal flank affects the conformation of the resulting assemblies. Significance: We identify the surface interfaces between Hsc70 and huntingtin exon 1, which allows the design of future therapeutic tools.

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“…Scale bar, 20 m. bitrap and both MS and MS/MS data analysis, as described under "Experimental Procedures" and in Ref. 29. Seven Hsc70-HttEx1Q25 cross-links were identified (Table 2).…”

Section: Hsc70 Effects On Httex1qn Assembly Are Independent Ofmentioning

confidence: 99%

Section: Hsc70 Effects On Httex1qn Assembly Are Independent Ofmentioning

confidence: 99%

Section: Expression and Purification Of Recombinant Polypeptides And mentioning

confidence: 99%

Section: Expression and Purification Of Recombinant Polypeptides And mentioning

confidence: 99%

See 2 more Smart Citations

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Monsellier

Redeker

Ruiz-Arlandis

et al. 2015

Journal of Biological Chemistry

Self Cite

106

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show abstract

“…BiP (HSPA5) AD/ Aβ Bound APP and reduced Aβ secretion [72]; reduced polyQ aggregation and toxicity in cells [73]; reduced α-synuclein toxicity in rat [74] and toxicity in cells and mice [79], ATPase mutant reduced large polyQ aggregates but no effect on toxicity [80], reduced axonal transport defect in polyQ fly [81]; binds α-synuclein and reduced toxicity of fibrils [82] [83]; binding to mutant SOD1 [84].…”

Section: Chaperone Family Chaperone Disease/protein(s) Commentsmentioning

confidence: 99%

Molecular chaperones and neuronal proteostasis

Smith¹,

Li²,

Cheetham³

2015

Seminars in Cell & Developmental Biology

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Protein homeostasis (proteostasis) is essential for maintaining the functionality of the proteome. The disruption of proteostasis, due to genetic mutations or an age-related decline, leads to aberrantly folded proteins that typically lose their function. The accumulation of misfolded and aggregated protein is also cytotoxic and has been implicated in the pathogenesis of neurodegenerative diseases. Neurons have developed an intrinsic protein quality control network, of which molecular chaperones are an essential component. Molecular chaperones function to promote efficient folding and target misfolded proteins for refolding or degradation. Increasing molecular chaperone expression can suppress protein aggregation and toxicity in numerous models of neurodegenerative disease; therefore, molecular chaperones are considered exciting therapeutic targets. Furthermore, mutations in several chaperones cause inherited neurodegenerative diseases. In this review, we focus on the importance of molecular chaperones in neurodegenerative diseases, and discuss the advances in understanding their protective mechanisms.

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Identification of protein interfaces within the multi‐aminoacyl‐tRNA synthetase complex: the case of lysyl‐tRNA synthetase and the scaffold protein p38

et al. 2016

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Human cytoplasmic lysyl‐ tRNA synthetase (Lys RS ) is associated within a multi‐aminoacyl‐ tRNA synthetase complex ( MSC ). Within this complex, the p38 component is the scaffold protein that binds the catalytic domain of Lys RS via its N‐terminal region. In addition to its translational function when associated to the MSC , Lys RS is also recruited in nontranslational roles after dissociation from the MSC . The balance between its MSC ‐associated and MSC ‐dissociated states is essential to regulate the functions of Lys RS in cellular homeostasis. With the aim of understanding the rules that govern association of Lys RS in the MSC , we analyzed the protein interfaces between Lys RS and the full‐length version of p38, the scaffold protein of the MSC . In a previous study, the cocrystal structure of Lys RS with a N‐terminal peptide of p38 was reported [Ofir‐Birin Y et al . (2013) Mol Cell 49, 30–42]. In order to identify amino acid residues involved in interaction of the two proteins, the non‐natural, photo‐cross‐linkable amino acid p ‐benzoyl‐ l ‐phenylalanine (Bpa) was incorporated at 27 discrete positions within the catalytic domain of Lys RS . Among the 27 distinct Lys RS mutants, only those with Bpa inserted in place of Lys356 or His364 were cross‐linked with p38. Using mass spectrometry, we unambiguously identified the protein interface of the cross‐linked complex and showed that Lys356 and His364 of Lys RS interact with the peptide from Pro8 to Arg26 in native p38, in agreement with the published cocrystal structure. This interface, which in Lys RS is located on the opposite side of the dimer to the site of interaction with its tRNA substrate, defines the core region of the MSC . The residues identified herein in human Lys RS are not conserved in yeast Lys RS , an enzyme that does not associate within the MSC , and contrast with the residues proposed to be essential for Lys RS :p38 association in the earlier work.

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Identification of Protein Interfaces between α-Synuclein, the Principal Component of Lewy Bodies in Parkinson Disease, and the Molecular Chaperones Human Hsc70 and the Yeast Ssa1p

Cited by 45 publications

References 42 publications

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Molecular Interaction between the Chaperone Hsc70 and the N-terminal Flank of Huntingtin Exon 1 Modulates Aggregation

Molecular chaperones and neuronal proteostasis

Identification of protein interfaces within the multi‐aminoacyl‐tRNA synthetase complex: the case of lysyl‐tRNA synthetase and the scaffold protein p38

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