Rapid Structural, Kinetic, and Immunochemical Analysis of Alpha-Synuclein Oligomers in Solution
Oligomers comprised of misfolded proteins are implicated as neurotoxins in the pathogenesis of protein misfolding conditions such as Parkinson’s and Alzheimer’s diseases. Structural, biophysical, and biochemical characterization of these nanoscale protein assemblies is key to understanding their pat...
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| Veröffentlicht in: | Nano letters 2020-11, Vol.20 (11), p.8163-8169 |
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| container_title | Nano letters |
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| creator | Arter, William E Xu, Catherine K Castellana-Cruz, Marta Herling, Therese W Krainer, Georg Saar, Kadi L Kumita, Janet R Dobson, Christopher M Knowles, Tuomas P. J |
| description | Oligomers comprised of misfolded proteins are implicated as neurotoxins in the pathogenesis of protein misfolding conditions such as Parkinson’s and Alzheimer’s diseases. Structural, biophysical, and biochemical characterization of these nanoscale protein assemblies is key to understanding their pathology and the design of therapeutic interventions, yet it is challenging due to their heterogeneous, transient nature and low relative abundance in complex mixtures. Here, we demonstrate separation of heterogeneous populations of oligomeric α-synuclein, a protein central to the pathology of Parkinson’s disease, in solution using microfluidic free-flow electrophoresis. We characterize nanoscale structural heterogeneity of transient oligomers on a time scale of seconds, at least 2 orders of magnitude faster than conventional techniques. Furthermore, we utilize our platform to analyze oligomer ζ-potential and probe the immunochemistry of wild-type α-synuclein oligomers. Our findings contribute to an improved characterization of α-synuclein oligomers and demonstrate the application of microchip electrophoresis for the free-solution analysis of biological nanoparticle analytes. |
| doi_str_mv | 10.1021/acs.nanolett.0c03260 |
| format | Article |
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J</creator><creatorcontrib>Arter, William E ; Xu, Catherine K ; Castellana-Cruz, Marta ; Herling, Therese W ; Krainer, Georg ; Saar, Kadi L ; Kumita, Janet R ; Dobson, Christopher M ; Knowles, Tuomas P. J</creatorcontrib><description>Oligomers comprised of misfolded proteins are implicated as neurotoxins in the pathogenesis of protein misfolding conditions such as Parkinson’s and Alzheimer’s diseases. Structural, biophysical, and biochemical characterization of these nanoscale protein assemblies is key to understanding their pathology and the design of therapeutic interventions, yet it is challenging due to their heterogeneous, transient nature and low relative abundance in complex mixtures. Here, we demonstrate separation of heterogeneous populations of oligomeric α-synuclein, a protein central to the pathology of Parkinson’s disease, in solution using microfluidic free-flow electrophoresis. We characterize nanoscale structural heterogeneity of transient oligomers on a time scale of seconds, at least 2 orders of magnitude faster than conventional techniques. Furthermore, we utilize our platform to analyze oligomer ζ-potential and probe the immunochemistry of wild-type α-synuclein oligomers. 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J</creatorcontrib><title>Rapid Structural, Kinetic, and Immunochemical Analysis of Alpha-Synuclein Oligomers in Solution</title><title>Nano letters</title><addtitle>NANO LETT</addtitle><addtitle>Nano Lett</addtitle><description>Oligomers comprised of misfolded proteins are implicated as neurotoxins in the pathogenesis of protein misfolding conditions such as Parkinson’s and Alzheimer’s diseases. Structural, biophysical, and biochemical characterization of these nanoscale protein assemblies is key to understanding their pathology and the design of therapeutic interventions, yet it is challenging due to their heterogeneous, transient nature and low relative abundance in complex mixtures. Here, we demonstrate separation of heterogeneous populations of oligomeric α-synuclein, a protein central to the pathology of Parkinson’s disease, in solution using microfluidic free-flow electrophoresis. We characterize nanoscale structural heterogeneity of transient oligomers on a time scale of seconds, at least 2 orders of magnitude faster than conventional techniques. Furthermore, we utilize our platform to analyze oligomer ζ-potential and probe the immunochemistry of wild-type α-synuclein oligomers. Our findings contribute to an improved characterization of α-synuclein oligomers and demonstrate the application of microchip electrophoresis for the free-solution analysis of biological nanoparticle analytes.</description><subject>alpha-Synuclein</subject><subject>Alzheimer Disease</subject><subject>Chemistry</subject><subject>Chemistry, Multidisciplinary</subject><subject>Chemistry, Physical</subject><subject>Humans</subject><subject>Materials Science</subject><subject>Materials Science, Multidisciplinary</subject><subject>Nanoscience & Nanotechnology</subject><subject>Parkinson Disease</subject><subject>Physical Sciences</subject><subject>Physics</subject><subject>Physics, Applied</subject><subject>Physics, Condensed Matter</subject><subject>Science & Technology</subject><subject>Science & Technology - Other Topics</subject><subject>Technology</subject><issn>1530-6984</issn><issn>1530-6992</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2020</creationdate><recordtype>article</recordtype><sourceid>AOWDO</sourceid><sourceid>EIF</sourceid><recordid>eNqNkctKAzEUhoMoVqtvIJK9nZpMMpdshFK8FAXB6jpkMhmbkknKJKP07U1pLboRVznh_P-5fAeAC4zGGKX4Wkg_tsI6o0IYI4lImqMDcIIzgpKcsfRwH5d0AE69XyKEGMnQMRgQggqWZeQE8Bex0jWch66Xoe-EGcFHbVXQcgSFreGsbXvr5EK1WgoDJ1aYtdceugZOzGohkvna9tIobeGz0e-uVZ2H8TN3pg_a2TNw1Ajj1fnuHYK3u9vX6UPy9Hw_m06eEkEpC0mF8rRhmBSFyBpJ0qyMI7JKlIUqKMsKUpMCkVpViDKaqpyUVOR5ShvCCKZlTYbgZlt31VetqqWyIS7DV51uRbfmTmj-O2P1gr-7D15gnJexwRDQbQHZOe871ey9GPENcB6B82_gfAc82i5_9t2bvglHQbkVfKrKNV5qZaXay-JJMpZuNowRZVMdxIba1PU2ROvV_61RjbbqzZxL13fxVP7v4b8Ass-yRQ</recordid><startdate>20201111</startdate><enddate>20201111</enddate><creator>Arter, William E</creator><creator>Xu, Catherine K</creator><creator>Castellana-Cruz, Marta</creator><creator>Herling, Therese W</creator><creator>Krainer, Georg</creator><creator>Saar, Kadi L</creator><creator>Kumita, Janet R</creator><creator>Dobson, Christopher M</creator><creator>Knowles, Tuomas P. J</creator><general>American Chemical Society</general><general>Amer Chemical Soc</general><scope>AOWDO</scope><scope>BLEPL</scope><scope>DTL</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>5PM</scope><orcidid>https://orcid.org/0000-0003-4726-636X</orcidid><orcidid>https://orcid.org/0000-0002-3887-4964</orcidid><orcidid>https://orcid.org/0000-0002-3615-1885</orcidid><orcidid>https://orcid.org/0000-0002-5926-3628</orcidid><orcidid>https://orcid.org/0000-0002-7879-0140</orcidid><orcidid>https://orcid.org/0000-0002-4726-636X</orcidid></search><sort><creationdate>20201111</creationdate><title>Rapid Structural, Kinetic, and Immunochemical Analysis of Alpha-Synuclein Oligomers in Solution</title><author>Arter, William E ; Xu, Catherine K ; Castellana-Cruz, Marta ; Herling, Therese W ; Krainer, Georg ; Saar, Kadi L ; Kumita, Janet R ; Dobson, Christopher M ; Knowles, Tuomas P. 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Here, we demonstrate separation of heterogeneous populations of oligomeric α-synuclein, a protein central to the pathology of Parkinson’s disease, in solution using microfluidic free-flow electrophoresis. We characterize nanoscale structural heterogeneity of transient oligomers on a time scale of seconds, at least 2 orders of magnitude faster than conventional techniques. Furthermore, we utilize our platform to analyze oligomer ζ-potential and probe the immunochemistry of wild-type α-synuclein oligomers. 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| subjects | alpha-Synuclein Alzheimer Disease Chemistry Chemistry, Multidisciplinary Chemistry, Physical Humans Materials Science Materials Science, Multidisciplinary Nanoscience & Nanotechnology Parkinson Disease Physical Sciences Physics Physics, Applied Physics, Condensed Matter Science & Technology Science & Technology - Other Topics Technology |
| title | Rapid Structural, Kinetic, and Immunochemical Analysis of Alpha-Synuclein Oligomers in Solution |
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