The Binding Mechanism of Epolactaene to Hsp60 Unveiled by in Silico Modelling

Spinello, Angelo; Barone, Giampaolo; Cappello, Francesco; Pace, Andrea; Buscemi, Silvestre; Piccionello, Antonio Palumbo

doi:10.1002/slct.201600125

Cited by 6 publications

(3 citation statements)

References 24 publications

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“…Epolactaene derivatives, such as the tertiary butyl ester ETB ( Figure 3 ), were also active [ 80 ] and SAR studies demonstrated that both the cyclic amide (lactam) and the α,β-unsaturated ketone are critical moieties for inhibiting the chaperone activity of Hsp60 [ 81 ]. An in-silico study suggested a putative mode of action for epolactaene, revealing the opening of a binding pocket in proximity of Cys442 after ATP occupies its binding site and that epolactaene covalently binds thiol moiety of Cys 442 through attack at C14 and epoxide ring-opening [ 82 ]. MD studies evidenced that epolactaene binding hinders the dynamic conformational changes of the monomer necessary for functional folding process.…”

Section: Hsp60mentioning

confidence: 99%

Heat Shock Proteins in Alzheimer’s Disease: Role and Targeting

Campanella

Pace

Bavisotto

et al. 2018

IJMS

Self Cite

133

104

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Among diseases whose cure is still far from being discovered, Alzheimer’s disease (AD) has been recognized as a crucial medical and social problem. A major issue in AD research is represented by the complexity of involved biochemical pathways, including the nature of protein misfolding, which results in the production of toxic species. Considering the involvement of (mis)folding processes in AD aetiology, targeting molecular chaperones represents a promising therapeutic perspective. This review analyses the connection between AD and molecular chaperones, with particular attention toward the most important heat shock proteins (HSPs) as representative components of the human chaperome: Hsp60, Hsp70 and Hsp90. The role of these proteins in AD is highlighted from a biological point of view. Pharmacological targeting of such HSPs with inhibitors or regulators is also discussed.

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Section: Hsp60mentioning

confidence: 99%

Heat Shock Proteins in Alzheimer’s Disease: Role and Targeting

Campanella

Pace

Bavisotto

et al. 2018

IJMS

Self Cite

133

104

View full text Add to dashboard Cite

show abstract

“…The transcriptional modifications, like S-guanylation in Cys 442 , can disturb the multichaperone complex stability and, therefore, alter the opening of the mitochondrial permeability pore 41 . In addition, covalent binding of metabolites like epolactaene or ETB (epolactaene tertiary butyl ester) to Cys 442 can induce the inhibition of HSP60 activity by disrupting oligomerization 28 or inducing conformational changes of the apical domain, responsible for the recognition of the co-chaperone HSP10 42 . In this sense, N-ethyl-maleimide or fluorescein-5maleimide modified the activity of HSP60 in steroidogenic cells by blocking cysteines through a thiol-maleimide reaction 27 that inhibits the protein activity by blocking cysteines 43 .…”

Section: Discussionmentioning

confidence: 99%

Characterization of Mitochondrial Heat Shock Protein 60 variants in HEK293 Cells Transformed into Steroidogenic

Arciga,

Sánchez,

Ménde

et al. 2023

MRAJ

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Introduction: During pregnancy, P4 is essential to maintain the maternal-fetal relationship. Maternal cholesterol is the main source of P4 production, a process that takes place in the syncytiotrophoblast mitochondrion. The mechanism and proteins involved in the cholesterol transport for the steroidogenic process are still unknown in detail. The STARD3 protein could be the substitute for its STARD1 equivalent localized in all acute response tissues. However, mutation or null STARD3 mice maintain their reproductive capacity, suggesting other proteins are involved in this process. Previously, we reported that the HSP60 participates in steroidogenesis in mitochondria isolated from the placental syncytiotrophoblast, mitochondrial contact sites or JEG-3. Also, take relevance that non-steroidogenic cells, such as the HEK293, which are human kidney embryo cells, when are transformed into steroidogenic by transfection of the steroidogenic machinery, they synthesize progesterone. To understand better the mechanism through which HSP60 participates in placental steroidogenesis, mutation of cysteine 442, which is essential in the active site for its activity, and deletion of 146 amino acid residues of the N-terminal of HSP60 were performed. The first was implemented to determine whether the protein structure is essential to support steroidogenesis, and the second was done to elucidate whether its activity occurs outside or inside the mitochondrion. Methods: Two mutants were obtained: a) cysteine 442 was replaced by alanine (HSP60C442A) and b) the HSP60-mature (HSP60M) without the mitochondrial-leading sequence. Human kidney cells HEK293 were transformed into steroidogenic by transfection with pECE-P450scc, pCMV-3βHSD-I. The transfected cells were transfected with the HSP60wt, HSP60C442A, or HSP60M plasmids. The transfection was validated by western blot and P4 was determined by an enzyme immunoassay kit. HSP60 without mutations was used as control (HSP60wt). Results: The synthesis of P4 was stimulated by the wild type HSP60 (HSP60wt). However, with both mutants, steroidogenesis occurred as in the control, suggesting that mutants do not support P4 synthesis. Discussion: The mechanism to transport cholesterol to steroidogenic mitochondria requires the full HSP60 to support P4 synthesis, which is necessary to maintain pregnancy. Highlights HSP60 participates in the steroidogenesis of transformed HEK293 cells. Cys442 mutant of HSP60 loses its activity in steroidogenesis. N-terminal deletion of HSP60 is not involved in steroidogenesis. Native HSP60 is critical for steroidogenesis.

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“…Owing to the fact that myrtucommulone has other biological targets and, in addition to Hsp60, affects the arachidonic acid metabolism, this compound can provide a starting point for the development of analogs deprived of these activities [229]. Epolactaene, found in the fungal strain Penicillium sp., is another natural product reported to selectively inhibit Hsp60 activity through covalent binding to cysteine residues close to the ATP binding pocket, which indicates the allosteric regulation of Hsp60 without affecting its ATPase activity [230]. The folding activity of the Hsp60/Hsp10 complex can also be disturbed by curcumin in a dose-dependent manner, as reported in neuroblastoma cells [231].…”

Section: Hsp60-aimed Therapiesmentioning

confidence: 91%

Heat Shock Proteins, a Double-Edged Sword: Significance in Cancer Progression, Chemotherapy Resistance and Novel Therapeutic Perspectives

Kunachowicz,

Król-Kulikowska,

Raczycka

et al. 2024

Cancers

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Heat shock proteins (Hsps) are involved in one of the adaptive mechanisms protecting cells against environmental and metabolic stress. Moreover, the large role of these proteins in the carcinogenesis process, as well as in chemoresistance, was noticed. This review aims to draw attention to the possibilities of using Hsps in developing new cancer therapy methods, as well as to indicate directions for future research on this topic. In order to discuss this matter, a thorough review of the latest scientific literature was carried out, taking into account the importance of selected proteins from the Hsp family, including Hsp27, Hsp40, Hsp60, Hsp70, Hsp90 and Hsp110. One of the more characteristic features of all Hsps is that they play a multifaceted role in cancer progression, which makes them an obvious target for modern anticancer therapy. Some researchers emphasize the importance of directly inhibiting the action of these proteins. In turn, others point to their possible use in the design of cancer vaccines, which would work by inducing an immune response in various types of cancer. Due to these possibilities, it is believed that the use of Hsps may contribute to the progress of oncoimmunology, and thus help in the development of modern anticancer therapies, which would be characterized by higher effectiveness and lower toxicity to the patients.

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The Binding Mechanism of Epolactaene to Hsp60 Unveiled by in Silico Modelling

Cited by 6 publications

References 24 publications

Heat Shock Proteins in Alzheimer’s Disease: Role and Targeting

Heat Shock Proteins in Alzheimer’s Disease: Role and Targeting

Characterization of Mitochondrial Heat Shock Protein 60 variants in HEK293 Cells Transformed into Steroidogenic

Heat Shock Proteins, a Double-Edged Sword: Significance in Cancer Progression, Chemotherapy Resistance and Novel Therapeutic Perspectives

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