Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants
Misfolding and aggregation of the neuronal protein alpha-synuclein (aSyn) has been identified as a hallmark of Parkinson’s disease (PD) pathology and other synucleinopathies. Preventing formation of intracellular aSyn accumulations constitutes a therapeutic strategy against disease development. We r...
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Frontiers Media S.A.
2025-05-01
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| author | Linnea Charlotta Hjelm Wojciech Paslawski Christofer Lendel Siri Flemming Svedmark Per Svenningsson Stefan Ståhl Hanna Lindberg John Löfblom |
| author_facet | Linnea Charlotta Hjelm Wojciech Paslawski Christofer Lendel Siri Flemming Svedmark Per Svenningsson Stefan Ståhl Hanna Lindberg John Löfblom |
| author_sort | Linnea Charlotta Hjelm |
| collection | DOAJ |
| description | Misfolding and aggregation of the neuronal protein alpha-synuclein (aSyn) has been identified as a hallmark of Parkinson’s disease (PD) pathology and other synucleinopathies. Preventing formation of intracellular aSyn accumulations constitutes a therapeutic strategy against disease development. We recently reported on a new type of affinity protein, denoted Sequestrin, aimed for efficient and stable interactions with aggregation-prone amyloidogenic proteins and peptides. Upon binding, sequestrins interact with the aggregation-prone peptide and form a stabilizing four-stranded beta sheet with similarities to the beta sheet rich structures seen in amyloid fibrils. Here, high-affinity aSyn-binding sequestrins were isolated from a large naïve sequestrin library using phage display technology. The best binders demonstrated dissociation constant, KD, values in the 10 nM-range, and structural rearrangements in both the sequestrin and aSyn protein upon binding. Modelling using AlphaFold, followed by NMR spectroscopy suggested that the sequestrins bind an N-terminal region of aSyn that is critical for amyloidogenic aggregation. In an in vitro aggregation study, the sequestrins demonstrated complete inhibition of aSyn aggregation at equimolar concentrations, including the three familial mutants A30P, E46K, and A53T that are associated with Parkinson’s disease and Lewy body dementia. |
| format | Article |
| id | doaj-art-2b6181bfb98b4ddb8d5df5dfa0083a4e |
| institution | Kabale University |
| issn | 1664-3224 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Frontiers Media S.A. |
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| series | Frontiers in Immunology |
| spelling | doaj-art-2b6181bfb98b4ddb8d5df5dfa0083a4e2025-08-20T03:49:46ZengFrontiers Media S.A.Frontiers in Immunology1664-32242025-05-011610.3389/fimmu.2025.15747551574755Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutantsLinnea Charlotta Hjelm0Wojciech Paslawski1Christofer Lendel2Siri Flemming Svedmark3Per Svenningsson4Stefan Ståhl5Hanna Lindberg6John Löfblom7Department of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, SwedenDepartment of Clinical Neuroscience, Karolinska Institute, Stockholm, SwedenDepartment of Chemistry, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, SwedenDepartment of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, SwedenDepartment of Clinical Neuroscience, Karolinska Institute, Stockholm, SwedenDepartment of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, SwedenDepartment of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, SwedenDepartment of Protein Science, School of Engineering Sciences in Chemistry, Biotechnology and Health, KTH Royal Institute of Technology, Stockholm, SwedenMisfolding and aggregation of the neuronal protein alpha-synuclein (aSyn) has been identified as a hallmark of Parkinson’s disease (PD) pathology and other synucleinopathies. Preventing formation of intracellular aSyn accumulations constitutes a therapeutic strategy against disease development. We recently reported on a new type of affinity protein, denoted Sequestrin, aimed for efficient and stable interactions with aggregation-prone amyloidogenic proteins and peptides. Upon binding, sequestrins interact with the aggregation-prone peptide and form a stabilizing four-stranded beta sheet with similarities to the beta sheet rich structures seen in amyloid fibrils. Here, high-affinity aSyn-binding sequestrins were isolated from a large naïve sequestrin library using phage display technology. The best binders demonstrated dissociation constant, KD, values in the 10 nM-range, and structural rearrangements in both the sequestrin and aSyn protein upon binding. Modelling using AlphaFold, followed by NMR spectroscopy suggested that the sequestrins bind an N-terminal region of aSyn that is critical for amyloidogenic aggregation. In an in vitro aggregation study, the sequestrins demonstrated complete inhibition of aSyn aggregation at equimolar concentrations, including the three familial mutants A30P, E46K, and A53T that are associated with Parkinson’s disease and Lewy body dementia.https://www.frontiersin.org/articles/10.3389/fimmu.2025.1574755/fullaffibody moleculealpha-synucleindirected evolutionParkinson’s diseasephage displaysequestrin |
| spellingShingle | Linnea Charlotta Hjelm Wojciech Paslawski Christofer Lendel Siri Flemming Svedmark Per Svenningsson Stefan Ståhl Hanna Lindberg John Löfblom Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants Frontiers in Immunology affibody molecule alpha-synuclein directed evolution Parkinson’s disease phage display sequestrin |
| title | Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants |
| title_full | Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants |
| title_fullStr | Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants |
| title_full_unstemmed | Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants |
| title_short | Engineered sequestrins inhibit aggregation of pathogenic alpha-synuclein mutants |
| title_sort | engineered sequestrins inhibit aggregation of pathogenic alpha synuclein mutants |
| topic | affibody molecule alpha-synuclein directed evolution Parkinson’s disease phage display sequestrin |
| url | https://www.frontiersin.org/articles/10.3389/fimmu.2025.1574755/full |
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