Modulating the Effects of the Bacterial Chaperonin GroEL on Fibrillogenic Polypeptides through Modification of Domain Hinge Architecture

Fukui, Naoya; Araki, Kiho; Hongo, Kunihiro; Mizobata, Tomohiro; Kawata, Yasushi

doi:10.1074/jbc.m116.751925

Cited by 15 publications

(12 citation statements)

References 36 publications

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“…We conclude that both Hsp60 WT and GW are stable tetradecamers, supporting a previous report [35]. Taken together with the results of GroEL G192W [24,30], we concluded that Hsp60 G190W had the open-mimic conformation of AD, similar to GroEL G192W, with hydrophobic characteristics that probably enhanced the amyloid fibrillation suppression effect. Comparing the structural characteristics of wild type (WT) and G190W Hsp60.…”

Section: Hsp60 Gw Shows An Increased Degree Of Surface Hydrophobicitysupporting

confidence: 88%

“…Our previous studies showed that the GroEL G192W mutant "locks" the GroEL oligomer in an open conformation through introduction of tryptophan at hinge II site pushes AD upward. This enhances the affinity of GroEL G192W toward fibrillogenic polypeptides [24,30]. We intended to apply this method to human Hsp60 (Figure 1a) in this study.…”

Section: Open-mimic Mutant Hsp60 G190w (Gw) Is a Potent Suppressor Ofmentioning

confidence: 99%

“…Previous studies have shown that the apical domain of the group I Hsp60 GroEL has a strong influence on α-synuclein amyloid fibrillation. We found that the apical domain (AD) open-mimic mutant GroEL, of which the glycine(G)192 at hinge II was mutated to tryptophan(W), has a powerful suppressive effect on amyloid fibril formation [24]. In addition, it has been shown that the isolated GroEL AD alone suppresses amyloid fibrillization of proteins such as α-synuclein and Aβ 1-42 , and functions as a mini-chaperone [25].…”

Section: Introductionmentioning

confidence: 99%

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Human Molecular Chaperone Hsp60 and Its Apical Domain Suppress Amyloid Fibril Formation of α-Synuclein

Yamamoto

Fukui

Adachi

et al. 2019

IJMS

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Heat shock proteins play roles in assisting other proteins to fold correctly and in preventing the aggregation and accumulation of proteins in misfolded conformations. However, the process of aging significantly degrades this ability to maintain protein homeostasis. Consequently, proteins with incorrect conformations are prone to aggregate and accumulate in cells, and this aberrant aggregation of misfolded proteins may trigger various neurodegenerative diseases, such as Parkinson’s disease. Here, we investigated the possibilities of suppressing α-synuclein aggregation by using a mutant form of human chaperonin Hsp60, and a derivative of the isolated apical domain of Hsp60 (Hsp60 AD(Cys)). In vitro measurements were used to detect the effects of chaperonin on amyloid fibril formation, and interactions between Hsp60 proteins and α-synuclein were probed by quartz crystal microbalance analysis. The ability of Hsp60 AD(Cys) to suppress α-synuclein intracellular aggregation and cytotoxicity was also demonstrated. We show that Hsp60 mutant and Hsp60 AD(Cys) both effectively suppress α-synuclein amyloid fibril formation, and also demonstrate for the first time the ability of Hsp60 AD(Cys) to function as a mini-chaperone inside cells. These results highlight the possibility of using Hsp60 AD as a method of prevention and treatment of neurodegenerative diseases.

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Section: Hsp60 Gw Shows An Increased Degree Of Surface Hydrophobicitysupporting

confidence: 88%

Section: Open-mimic Mutant Hsp60 G190w (Gw) Is a Potent Suppressor Ofmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Human Molecular Chaperone Hsp60 and Its Apical Domain Suppress Amyloid Fibril Formation of α-Synuclein

Yamamoto

Fukui

Adachi

et al. 2019

IJMS

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“…Furthermore, the abundance of chaperonin GroEL (spot 1058; p-value = 0.006), universal stress protein (spot 3811), and quinoprotein ethanol dehydrogenase (spot 1001; p-value = 0.010) increased significantly during the time-course of biopolymer synthesis. GroEL, as the chaperonin family of molecular chaperones, is involved in the proper folding of many proteins [29]. Therefore, its increased abundance during P. putida KT2440 growth was important for aggregated proteins to renature after exposure to unfavorable conditions.…”

Section: Stress Responsementioning

confidence: 99%

Proteomic Response of Pseudomonas putida KT2440 to Dual Carbon-Phosphorus Limitation during mcl-PHAs Synthesis

Możejko-Ciesielska

Serafim

2019

Biomolecules

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Pseudomonas putida KT2440, one of the best characterized pseudomonads, is a metabolically versatile producer of medium-chain-length polyhydroxyalkanoates (mcl-PHAs) that serves as a model bacterium for molecular studies. The synthesis of mcl-PHAs is of great interest due to their commercial potential. Carbon and phosphorus are the essential nutrients for growth and their limitation can trigger mcl-PHAs’ production in microorganisms. However, the specific molecular mechanisms that drive this synthesis in Pseudomonas species under unfavorable growth conditions remain poorly understood. Therefore, the proteomic responses of Pseudomonas putida KT2440 to the limited carbon and phosphorus levels in the different growth phases during mcl-PHAs synthesis were investigated. The data indicated that biopolymers’ production was associated with the cell growth of P. putida KT2440 under carbon- and phosphorus-limiting conditions. The protein expression pattern changed during mcl-PHAs synthesis and accumulation, and during the different physiological states of the microorganism. The data suggested that the majority of metabolic activities ceased under carbon and phosphorus limitation. The abundance of polyhydroxyalkanoate granule-associated protein (PhaF) involved in PHA synthesis increased significantly at 24 and 48 h of the cultivations. The activation of proteins belonging to the phosphate regulon was also detected. Moreover, these results indicated changes in the protein profiles related to amino acids metabolism, replication, transcription, translation, stress response mechanisms, transport or signal transduction. The presented data allowed the investigation of time-course proteome alterations in response to carbon and phosphorus limitation, and PHAs synthesis. This study provided information about proteins that can be potential targets in improving the efficiency of mcl-PHAs synthesis.

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“…In an effort analogous to our previous studies to use bacterial chaperones for protein aggregation control (14,15), we set out to probe the possibilities of using HdeA and HdeB in the control of irreversible aggregation and fibrillation of proteins, specifically at low pH. We began our experiments using human ␣-Synuclein because previous studies by Uversky et al (16) demonstrated that this protein forms fibrils more readily at lower pH (pH Ͻ5) than at neutral pH ( Fig.…”

mentioning

confidence: 99%

Acid-denatured small heat shock protein HdeA from Escherichia coli forms reversible fibrils with an atypical secondary structure

Miyawaki

Uemura

Hongo

et al. 2019

Journal of Biological Chemistry

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The periplasmic small heat shock protein HdeA from Escherichia coli is inactive under normal growth conditions (at pH 7) and activated only when E. coli cells are subjected to a sudden decrease in pH, converting HdeA into an acid-denatured active state. Here, using in vitro fibrillation assays, transmission EM, atomic-force microscopy, and CD analyses, we found that when HdeA is active as a molecular chaperone, it is also capable of forming inactive aggregates that, at first glance, resemble amyloid fibrils. We noted that the molecular chaperone activity of HdeA takes precedence over fibrillogenesis under acidic conditions, as the presence of denatured substrate protein was sufficient to suppress HdeA fibril formation. Further experiments suggested that the secondary structure of HdeA fibrils deviates somewhat from typical amyloid fibrils and contains α-helices. Strikingly, HdeA fibrils that formed at pH 2 were immediately resolubilized by a simple shift to pH 7 and from there could regain molecular chaperone activity upon a return to pH 1. HdeA, therefore, provides an unusual example of a “reversible” form of protein fibrillation with an atypical secondary structure composition. The competition between active assistance of denatured polypeptides (its “molecular chaperone” activity) and the formation of inactive fibrillary deposits (its “fibrillogenic” activity) provides a unique opportunity to probe the relationship among protein function, structure, and aggregation in detail.

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Modulating the Effects of the Bacterial Chaperonin GroEL on Fibrillogenic Polypeptides through Modification of Domain Hinge Architecture

Cited by 15 publications

References 36 publications

Human Molecular Chaperone Hsp60 and Its Apical Domain Suppress Amyloid Fibril Formation of α-Synuclein

Human Molecular Chaperone Hsp60 and Its Apical Domain Suppress Amyloid Fibril Formation of α-Synuclein

Proteomic Response of Pseudomonas putida KT2440 to Dual Carbon-Phosphorus Limitation during mcl-PHAs Synthesis

Acid-denatured small heat shock protein HdeA from Escherichia coli forms reversible fibrils with an atypical secondary structure

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