Fibril Formation of hsp10 Homologue Proteins and Determination of Fibril Core Regions: Differences in Fibril Core Regions Dependent on Subtle Differences in Amino Acid Sequence

Yagi, Hisashi; Sato, Akira; Yoshida, Akihiro; Hattori, Yoshiki; Hara, Masahiro; Shimamura, Jun; Sakane, Isao; Hongo, Kunihiro; Mizobata, Tomohiro; Kawata, Yasushi

doi:10.1016/j.jmb.2008.02.012

Cited by 11 publications

(18 citation statements)

References 30 publications

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“…We have previously found that GroES may form typical amyloid fibrils from the guanidine hydrochloride (Gdn-HCl) 2 unfolded state (19) and determined the core sequence of the mature GroES amyloid fibrils that are resistant to protease (Fig. 1B) (20). Our findings proved that even a molecular chaperone may form amyloid fibrils under certain conditions, supporting the recent finding that amyloid fibrils are an inherently common structure of many proteins under certain conditions in vitro (21)(22)(23).…”

supporting

confidence: 84%

“…Interestingly, this region was adjacent to the fibril core sequence that we determined previously (20). It was also found that formation of this intermediate hinged on an Asn-Gly rearrangement reaction, which yielded ␤-aspartic acid as a consequence of deamidation of the asparagine.…”

supporting

confidence: 55%

“…Because after these changes formation of amyloid fibrils was confirmed by Thioflavin-T binding assay in separate experiments, these data strongly suggested the appearance of soluble intermediate species formed during nucleus formation. These residues were adjacent to the fibril core region (Asp 58 -Lys 74 ) previously identified (20) in GroES amyloid fibrils.…”

Section: Detection Of Structural Changes From the Disordered Stateto mentioning

confidence: 74%

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Covalent Structural Changes in Unfolded GroES That Lead to Amyloid Fibril Formation Detected by NMR

Iwasa

Meshitsuka

Hongo

et al. 2011

Journal of Biological Chemistry

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Co-chaperonin GroES from Escherichia coli works with chaperonin GroEL to mediate the folding reactions of various proteins. However, under specific conditions, i.e. the completely disordered state in guanidine hydrochloride, this molecular chaperone forms amyloid fibrils similar to those observed in various neurodegenerative diseases. Thus, this is a good model system to understand the amyloid fibril formation mechanism of intrinsically disordered proteins. Here, we identified a critical intermediate of GroES in the early stages of this fibril formation using NMR and mass spectroscopy measurements. A covalent rearrangement of the polypeptide bond at Asn 45 -Gly 46 and/or Asn 51 -Gly 52 that eventually yield ␤-aspartic acids via deamidation of asparagine was observed to precede fibril formation. Mutation of these asparagines to alanines resulted in delayed nucleus formation. Our results indicate that peptide bond rearrangement at Asn-Gly enhances the formation of GroES amyloid fibrils. The finding provides a novel insight into the structural process of amyloid fibril formation from a disordered state, which may be applicable to intrinsically disordered proteins in general.Intrinsically disordered proteins are commonly defined as proteins that do not adopt a well defined structure in solution (1), and they can fold into ordered structures only upon binding to their cellular targets (2). They are abundant in eukaryotic proteins and play a significant role in biological functions associated with signaling and regulation events (3). Some intrinsically disordered proteins, exemplified by ␣-synuclein (4 -6) and poly(Q) (7-9) proteins, are capable of forming insoluble aggregates referred to as amyloid fibrils. Understanding the detailed mechanisms in which intrinsically disordered proteins self-assemble into amyloid fibrils is a very important issue because they associate closely with amyloid-related degenerative diseases (10).The prevalent model to explain the mechanisms of amyloid fibril formation in vitro is nucleation-dependent fibril formation (11). Fibril formation kinetics consists of two phases, that is, nucleation and extension, as traced with Thioflavin-T-binding fluorescence or turbidity of incubated samples. Nucleus formation requires a series of associations between protein monomers that are thermodynamically unfavorable, and this step represents the rate-limiting step in amyloid fibril formation. Once the nucleus has been formed, the further addition of monomers becomes thermodynamically favorable, resulting in a rapid extension of amyloid fibrils (12, 13). It has generally been assumed until recently that the cytotoxicity in amyloidrelated degenerative diseases was due to mature amyloid fibrils, but now attention has shifted to various intermediate species formed during nucleation, such as an oligomeric but soluble state (14 -16) or even some monomeric states (17, 18). Therefore, details regarding the structural characteristics of these species that appear in the early stage of the fibril formation are ve...

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supporting

confidence: 84%

supporting

confidence: 55%

Section: Detection Of Structural Changes From the Disordered Stateto mentioning

confidence: 74%

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Covalent Structural Changes in Unfolded GroES That Lead to Amyloid Fibril Formation Detected by NMR

Iwasa

Meshitsuka

Hongo

et al. 2011

Journal of Biological Chemistry

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“…4. This peak also contained intact and 20 and Peak 5 was found to be a mixture of two peptides, 1 Ala-Thr-Lys-Ala-Val-Cys-ValLeu-Lys-Gly-Asp-Gly-Pro-Val-Gln-Gly-Ile-Ile-AsnPhe-Glu-Gln-Lys-Glu-Ser-Asn-Gly-Pro-Val-Lys-ValTrp 32 and 89 Ala-Asp-Lys-Asp-Gly-Val-Ala-Asp-ValSer-Ile-Glu-Asp-Ser-Val-Ile-Ser-Leu-Ser-Gly-Asp-HisCys-Ile-Ile-Gly-Arg-Thr-Leu-Val-Val-His-Glu-Lys-Al a-Asp-Asp-Leu-Gly-Lys-Gly-Gly 130 . Of note was the identification of peptide (Gly-Ile-Ile-Asn-Phe) that was relatively short in length as a participant in fibril core formation.…”

Section: Determination Of Fibril Core Regionmentioning

confidence: 99%

“…Fibril core regions in SOD1 were determined according to the method described previously (32,33). Briefly, isolated precipitates (fibril core region) representing fractions of amyloid fibrils that had avoided digestion by A.lyticus protease I (API, Wako) or proteinase K (PK, Roche) were dissolved in 7.5 M Gdn-HCl, and individual peptides were isolated using reversed-phase high performance liquid chromatography (HPLC) on a CAPCELL PAK C-18 reverse phase column (Shiseido) and a Gilson HPLC system.…”

Section: Determination Of Amyloid Fibril Core Regionmentioning

confidence: 99%

Structural basis of Cu, Zn-superoxide dismutase amyloid fibril formation involves interaction of multiple peptide core regions

Ida¹,

Ando²,

Adachi³

et al. 2015

J Biochem

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Cu, Zn-superoxide dismutase (SOD1), an enzyme implicated in the progression of familial amyotrophic lateral sclerosis (fALS), forms amyloid fibrils under certain experimental conditions. As part of our efforts to understand ALS pathogenesis, in this study we found that reduction of the intramolecular disulfide bond destabilized the tertiary structure of metal free wildtype SOD1 and greatly enhanced fibril formation in vitro. We also identified fibril core peptides that are resistant to protease digestion by using mass spectroscopy and Edman degradation analyses. Three regions dispersed throughout the sequence were detected as fibril core sequences of SOD1. Interestingly, by using three synthetic peptides that correspond to these identified regions, we determined that each region was capable of fibril formation, either alone or in a mixture containing multiple peptides. It was also revealed that by reducing the disulfide bond and causing a decrease in the structural stability, the amyloid fibril formation of a familial mutant SOD1 G93A was accelerated even under physiological conditions. These results demonstrate that by destabilizing the structure of SOD1 by removing metal ions and breaking the intramolecular disulfide bridge, multiple fibril-forming core regions are exposed, which then interact with each another and form amyloid fibrils under physiological conditions.

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Role of C‐terminal negative charges and tyrosine residues in fibril formation of α‐synuclein

et al. 2012

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α-Synuclein (140 amino acids), one of the causative proteins of Parkinson's disease, forms amyloid fibrils in brain neuronal cells. In order to further explore the contributions of the C-terminal region of α-synuclein in fibril formation and also to understand the overall mechanism of fibril formation, we reduced the number of negatively charged residues in the C-terminal region using mutagenesis. Mutants with negative charges deleted displayed accelerated fibril formation compared with wild-type α-synuclein, demonstrating that negative charges located in the C-terminal region of α-synuclein modulate fibril formation. Additionally, when tyrosine residues located at position 125, 133, and 136 in the C-terminal region were changed to alanine residue(s), we found that all mutants containing the Tyr136Ala mutation showed delays in fibril formation compared with wild type. Mutation of Tyr136 to various amino acids revealed that aromatic residues located at this position act favorably toward fibril formation. In mutants where charge neutralization and tyrosine substitution were combined, we found that these two factors influence fibril formation in complex fashion. These findings highlight the importance of negative charges and aromatic side chains in the C-terminal region of α-synuclein in fibril formation.

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Fibril Formation of hsp10 Homologue Proteins and Determination of Fibril Core Regions: Differences in Fibril Core Regions Dependent on Subtle Differences in Amino Acid Sequence

Cited by 11 publications

References 30 publications

Covalent Structural Changes in Unfolded GroES That Lead to Amyloid Fibril Formation Detected by NMR

Covalent Structural Changes in Unfolded GroES That Lead to Amyloid Fibril Formation Detected by NMR

Structural basis of Cu, Zn-superoxide dismutase amyloid fibril formation involves interaction of multiple peptide core regions

Role of C‐terminal negative charges and tyrosine residues in fibril formation of α‐synuclein

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