Insight Into Factors Influencing the Aggregation Process in Wild‐Type and P66R Mutant SOD1 : Computational and Spectroscopic Approaches
Fuente:
Wiley Open Access
Salvato in:
| Autori principali: | , , , |
|---|---|
| Natura: | Artículo Open Access |
| Pubblicazione: |
Wiley
2024
|
| Soggetti: | |
| Accesso online: | |
| Tags: |
Aggiungi Tag
Nessun Tag, puoi essere il primo ad aggiungerne!!
|
| _version_ | 1867011970469724160 |
|---|---|
| author | Roghayeh Farrokhzad Bagher Seyedalipour Payam Baziyar Saman Hosseinkhani |
| author_facet | Roghayeh Farrokhzad Bagher Seyedalipour Payam Baziyar Saman Hosseinkhani Roghayeh Farrokhzad Bagher Seyedalipour Payam Baziyar Saman Hosseinkhani |
| collection | Wiley Open Access |
| contents | Insight Into Factors Influencing the Aggregation Process in Wild‐Type and P66R Mutant SOD1 : Computational and Spectroscopic Approaches Roghayeh Farrokhzad Bagher Seyedalipour Payam Baziyar Saman Hosseinkhani Proteins: Structure, Function, and Bioinformatics ABSTRACT Disturbances in metal ion homeostasis associated with amyotrophic lateral sclerosis (ALS) have been described for several years, but the exact mechanism of involvement is not well understood. To elucidate the role of metalation in superoxide dismutase (SOD1) misfolding and aggregation, we comprehensively characterized the structural features (apo/holo forms) of WT‐SOD1 and P66R mutant in loop IV. Using computational and experimental methodologies, we assessed the physicochemical properties of these variants and their correlation with protein aggregation at the molecular level. Modifications in apo‐SOD1 compared to holo‐SOD1 were more pronounced in flexibility, stability, hydrophobicity, and intramolecular interactions, as indicated by molecular dynamics simulations. The enzymatic activities of holo/apo‐WT SOD1 were 1.30 and 1.88‐fold of the holo/apo P66R mutant, respectively. Under amyloid‐inducing conditions, decreased ANS fluorescence intensity in the apo‐form relative to the holo‐form suggested pre‐fibrillar species and amyloid aggregate growth due to occluded hydrophobic pockets. FTIR spectroscopy revealed that apo‐WT‐SOD1 and apo‐P66R exhibited a mixture of parallel and intermolecular β‐sheet structures, indicative of aggregation propensity. Aggregate species were identified using TEM, Congo red staining, and ThT/ANS fluorescence spectroscopy. Thermodynamic analyses with GdnHCl demonstrated that metal deficit, mutation, and intramolecular disulfide bond reduction are essential for initiating SOD1 misfolding and aggregation. These disruptions destabilize the dimer‐monomer equilibrium, promoting dimer dissociation into monomers and decreasing the thermodynamic stability of SOD1 variants, thus facilitating amyloid/amorphous aggregate formation. Our findings offer novel insights into protein aggregation mechanisms in disease pathology and highlight potential therapeutic strategies against toxic protein aggregation, including SOD1. 10.1002/prot.26765 http://onlinelibrary.wiley.com/termsAndConditions#vor |
| doi_str_mv | 10.1002/prot.26765 |
| format | Artículo Open Access |
| id | wiley_oa_10_1002_prot_26765 |
| institution | Wiley Open Access |
| license_str_mv | http://onlinelibrary.wiley.com/termsAndConditions#vor |
| publishDate | 2024 |
| publisher | Wiley |
| record_format | wiley_oa |
| spellingShingle | Insight Into Factors Influencing the Aggregation Process in Wild‐Type and P66R Mutant SOD1 : Computational and Spectroscopic Approaches Roghayeh Farrokhzad Bagher Seyedalipour Payam Baziyar Saman Hosseinkhani Proteins: Structure, Function, and Bioinformatics Insight Into Factors Influencing the Aggregation Process in Wild‐Type and P66R Mutant SOD1 : Computational and Spectroscopic Approaches Roghayeh Farrokhzad Bagher Seyedalipour Payam Baziyar Saman Hosseinkhani Proteins: Structure, Function, and Bioinformatics ABSTRACT Disturbances in metal ion homeostasis associated with amyotrophic lateral sclerosis (ALS) have been described for several years, but the exact mechanism of involvement is not well understood. To elucidate the role of metalation in superoxide dismutase (SOD1) misfolding and aggregation, we comprehensively characterized the structural features (apo/holo forms) of WT‐SOD1 and P66R mutant in loop IV. Using computational and experimental methodologies, we assessed the physicochemical properties of these variants and their correlation with protein aggregation at the molecular level. Modifications in apo‐SOD1 compared to holo‐SOD1 were more pronounced in flexibility, stability, hydrophobicity, and intramolecular interactions, as indicated by molecular dynamics simulations. The enzymatic activities of holo/apo‐WT SOD1 were 1.30 and 1.88‐fold of the holo/apo P66R mutant, respectively. Under amyloid‐inducing conditions, decreased ANS fluorescence intensity in the apo‐form relative to the holo‐form suggested pre‐fibrillar species and amyloid aggregate growth due to occluded hydrophobic pockets. FTIR spectroscopy revealed that apo‐WT‐SOD1 and apo‐P66R exhibited a mixture of parallel and intermolecular β‐sheet structures, indicative of aggregation propensity. Aggregate species were identified using TEM, Congo red staining, and ThT/ANS fluorescence spectroscopy. Thermodynamic analyses with GdnHCl demonstrated that metal deficit, mutation, and intramolecular disulfide bond reduction are essential for initiating SOD1 misfolding and aggregation. These disruptions destabilize the dimer‐monomer equilibrium, promoting dimer dissociation into monomers and decreasing the thermodynamic stability of SOD1 variants, thus facilitating amyloid/amorphous aggregate formation. Our findings offer novel insights into protein aggregation mechanisms in disease pathology and highlight potential therapeutic strategies against toxic protein aggregation, including SOD1. 10.1002/prot.26765 http://onlinelibrary.wiley.com/termsAndConditions#vor |
| title | Insight Into Factors Influencing the Aggregation Process in Wild‐Type and P66R Mutant SOD1 : Computational and Spectroscopic Approaches |
| topic | Proteins: Structure, Function, and Bioinformatics |
| url | https://onlinelibrary.wiley.com/doi/10.1002/prot.26765 |