Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization
Alzheimer’s disease is the leading cause of dementia in the elderly. Pathologically it is characterized by the presence of amyloid plaques and neuronal loss within the brain tissue of affected individuals. It is now widely hypothesised that fibrillar structures represent an inert structure. Biophysi...
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PeerJ Inc.
2013-05-01
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| Online Access: | https://peerj.com/articles/73.pdf |
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| author | Timothy M. Ryan Joanne Caine Haydyn D.T. Mertens Nigel Kirby Julie Nigro Kerry Breheney Lynne J. Waddington Victor A. Streltsov Cyril Curtain Colin L. Masters Blaine R. Roberts |
| author_facet | Timothy M. Ryan Joanne Caine Haydyn D.T. Mertens Nigel Kirby Julie Nigro Kerry Breheney Lynne J. Waddington Victor A. Streltsov Cyril Curtain Colin L. Masters Blaine R. Roberts |
| author_sort | Timothy M. Ryan |
| collection | DOAJ |
| description | Alzheimer’s disease is the leading cause of dementia in the elderly. Pathologically it is characterized by the presence of amyloid plaques and neuronal loss within the brain tissue of affected individuals. It is now widely hypothesised that fibrillar structures represent an inert structure. Biophysical and toxicity assays attempting to characterize the formation of both the fibrillar and the intermediate oligomeric structures of Aβ typically involves preparing samples which are largely monomeric; the most common method by which this is achieved is to use the fluorinated organic solvent 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). Recent evidence has suggested that this method is not 100% effective in producing an aggregate free solution. We show, using dynamic light scattering, size exclusion chromatography and small angle X-ray scattering that this is indeed the case, with HFIP pretreated Aβ peptide solutions displaying an increased proportion of oligomeric and aggregated material and an increased propensity to aggregate. Furthermore we show that an alternative technique, involving treatment with strong alkali results in a much more homogenous solution that is largely monomeric. These techniques for solubilising and controlling the oligomeric state of Aβ are valuable starting points for future biophysical and toxicity assays. |
| first_indexed | 2024-03-09T06:51:14Z |
| format | Article |
| id | doaj.art-7b1183117385402f99febb60c89c705d |
| institution | Directory Open Access Journal |
| issn | 2167-8359 |
| language | English |
| last_indexed | 2024-03-09T06:51:14Z |
| publishDate | 2013-05-01 |
| publisher | PeerJ Inc. |
| record_format | Article |
| series | PeerJ |
| spelling | doaj.art-7b1183117385402f99febb60c89c705d2023-12-03T10:25:41ZengPeerJ Inc.PeerJ2167-83592013-05-011e7310.7717/peerj.7373Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterizationTimothy M. Ryan0Joanne Caine1Haydyn D.T. Mertens2Nigel Kirby3Julie Nigro4Kerry Breheney5Lynne J. Waddington6Victor A. Streltsov7Cyril Curtain8Colin L. Masters9Blaine R. Roberts10The Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, Victoria, AustraliaMaterials Science and Engineering, Preventative Health Flagship, Commonwealth Scientific and Industrial Research Organization, Parkville, Victoria, AustraliaSAXS/WAXS Beamline, Australian Synchrotron, Clayton, Victoria, AustraliaSAXS/WAXS Beamline, Australian Synchrotron, Clayton, Victoria, AustraliaMaterials Science and Engineering, Preventative Health Flagship, Commonwealth Scientific and Industrial Research Organization, Parkville, Victoria, AustraliaMaterials Science and Engineering, Preventative Health Flagship, Commonwealth Scientific and Industrial Research Organization, Parkville, Victoria, AustraliaMaterials Science and Engineering, Preventative Health Flagship, Commonwealth Scientific and Industrial Research Organization, Parkville, Victoria, AustraliaMaterials Science and Engineering, Preventative Health Flagship, Commonwealth Scientific and Industrial Research Organization, Parkville, Victoria, AustraliaThe Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, Victoria, AustraliaThe Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, Victoria, AustraliaThe Florey Institute of Neuroscience and Mental Health, University of Melbourne, Parkville, Victoria, AustraliaAlzheimer’s disease is the leading cause of dementia in the elderly. Pathologically it is characterized by the presence of amyloid plaques and neuronal loss within the brain tissue of affected individuals. It is now widely hypothesised that fibrillar structures represent an inert structure. Biophysical and toxicity assays attempting to characterize the formation of both the fibrillar and the intermediate oligomeric structures of Aβ typically involves preparing samples which are largely monomeric; the most common method by which this is achieved is to use the fluorinated organic solvent 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP). Recent evidence has suggested that this method is not 100% effective in producing an aggregate free solution. We show, using dynamic light scattering, size exclusion chromatography and small angle X-ray scattering that this is indeed the case, with HFIP pretreated Aβ peptide solutions displaying an increased proportion of oligomeric and aggregated material and an increased propensity to aggregate. Furthermore we show that an alternative technique, involving treatment with strong alkali results in a much more homogenous solution that is largely monomeric. These techniques for solubilising and controlling the oligomeric state of Aβ are valuable starting points for future biophysical and toxicity assays.https://peerj.com/articles/73.pdfAmyloidHexafluoroisopropanolAmmonium HydroxideAggregationA βA β toxicity |
| spellingShingle | Timothy M. Ryan Joanne Caine Haydyn D.T. Mertens Nigel Kirby Julie Nigro Kerry Breheney Lynne J. Waddington Victor A. Streltsov Cyril Curtain Colin L. Masters Blaine R. Roberts Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization PeerJ Amyloid Hexafluoroisopropanol Ammonium Hydroxide Aggregation A β A β toxicity |
| title | Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization |
| title_full | Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization |
| title_fullStr | Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization |
| title_full_unstemmed | Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization |
| title_short | Ammonium hydroxide treatment of Aβ produces an aggregate free solution suitable for biophysical and cell culture characterization |
| title_sort | ammonium hydroxide treatment of aβ produces an aggregate free solution suitable for biophysical and cell culture characterization |
| topic | Amyloid Hexafluoroisopropanol Ammonium Hydroxide Aggregation A β A β toxicity |
| url | https://peerj.com/articles/73.pdf |
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