Novel methods to study amyloid oligomers
Ortigosa Pascual, Lei LU
; Andrzejewska, Ewa A.
LU
; Pálmadóttir, Tinna
LU
; Knowles, Tuomas P.
and Linse, Sara
LU
(2024)
In Biophysical Journal
123(3).
p.41-41
- Abstract
- Amyloid protein deposits are the hallmark of neurodegenerative diseases such as Alzheimer’s, Parkinson’s, or diabetes type II. Although their connection to the diseases is not fully understood, many studies point at intermediate species called oligomers as being the main responsible for the toxicity of these proteins. This makes them strong candidates as therapeutic targets. However, due to their low relative concentration, heterogenicity and transient nature, methods to study and characterize oligomers are lacking. First, we optimized the cross-linking method photo-induced crosslinking of unmodified proteins (PICUP). By cross-linking monomers within an oligomer together, the otherwise transient interaction can be turned into covalent... (More)
- Amyloid protein deposits are the hallmark of neurodegenerative diseases such as Alzheimer’s, Parkinson’s, or diabetes type II. Although their connection to the diseases is not fully understood, many studies point at intermediate species called oligomers as being the main responsible for the toxicity of these proteins. This makes them strong candidates as therapeutic targets. However, due to their low relative concentration, heterogenicity and transient nature, methods to study and characterize oligomers are lacking. First, we optimized the cross-linking method photo-induced crosslinking of unmodified proteins (PICUP). By cross-linking monomers within an oligomer together, the otherwise transient interaction can be turned into covalent ones, allowing one to study oligomers with great ease. Our optimized method has allowed us to observe cross-linking of alpha-synuclein (αSyn) as quickly as with 1 ms reaction, and we have proven this method to be a valuable tool for oligomer studies. Moreover, we have developed a mass-spectrometry analysis protocol to gain structural insight into the oligomers, allowing us to understand the behavior of αSyn oligomers during aggregation, their binding to lipid vesicles, and their interaction with chaperone DNJB6. Second, we have made use of a custom made confocal fluorescence set-up to perform microfluidic free flow electrophoresis on amyloid-beta (Aβ42). This method allows us to separate oligomers of different electrophoretic mobility from each other and perform single-molecule fluorescence measurements to count the amount and size of every species in a single sample. With the help of this method, we have studied differences in oligomer population between idle and agitated aggregation, synthetic and recombinant Aβ42 and even the ability of fibrils to catalyze oligomer dissociation. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/6ef3c4ad-2dbf-43a5-8d4c-c14b6eb202bf
- author
- Ortigosa Pascual, Lei
LU
; Andrzejewska, Ewa A.
LU
; Pálmadóttir, Tinna
LU
; Knowles, Tuomas P.
and Linse, Sara
LU
- organization
- publishing date
- 2024-02-08
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Biophysical Journal
- volume
- 123
- issue
- 3
- pages
- 1 pages
- publisher
- Cell Press
- ISSN
- 1542-0086
- DOI
- 10.1016/j.bpj.2023.11.331
- language
- English
- LU publication?
- yes
- id
- 6ef3c4ad-2dbf-43a5-8d4c-c14b6eb202bf
- date added to LUP
- 2024-12-02 18:16:09
- date last changed
- 2025-04-04 15:16:12
@misc{6ef3c4ad-2dbf-43a5-8d4c-c14b6eb202bf,
abstract = {{Amyloid protein deposits are the hallmark of neurodegenerative diseases such as Alzheimer’s, Parkinson’s, or diabetes type II. Although their connection to the diseases is not fully understood, many studies point at intermediate species called oligomers as being the main responsible for the toxicity of these proteins. This makes them strong candidates as therapeutic targets. However, due to their low relative concentration, heterogenicity and transient nature, methods to study and characterize oligomers are lacking. First, we optimized the cross-linking method photo-induced crosslinking of unmodified proteins (PICUP). By cross-linking monomers within an oligomer together, the otherwise transient interaction can be turned into covalent ones, allowing one to study oligomers with great ease. Our optimized method has allowed us to observe cross-linking of alpha-synuclein (αSyn) as quickly as with 1 ms reaction, and we have proven this method to be a valuable tool for oligomer studies. Moreover, we have developed a mass-spectrometry analysis protocol to gain structural insight into the oligomers, allowing us to understand the behavior of αSyn oligomers during aggregation, their binding to lipid vesicles, and their interaction with chaperone DNJB6. Second, we have made use of a custom made confocal fluorescence set-up to perform microfluidic free flow electrophoresis on amyloid-beta (Aβ<sub>42</sub>). This method allows us to separate oligomers of different electrophoretic mobility from each other and perform single-molecule fluorescence measurements to count the amount and size of every species in a single sample. With the help of this method, we have studied differences in oligomer population between idle and agitated aggregation, synthetic and recombinant Aβ<sub>42</sub> and even the ability of fibrils to catalyze oligomer dissociation.}},
author = {{Ortigosa Pascual, Lei and Andrzejewska, Ewa A. and Pálmadóttir, Tinna and Knowles, Tuomas P. and Linse, Sara}},
issn = {{1542-0086}},
language = {{eng}},
month = {{02}},
note = {{Conference Abstract}},
number = {{3}},
pages = {{41--41}},
publisher = {{Cell Press}},
series = {{Biophysical Journal}},
title = {{Novel methods to study amyloid oligomers}},
url = {{http://dx.doi.org/10.1016/j.bpj.2023.11.331}},
doi = {{10.1016/j.bpj.2023.11.331}},
volume = {{123}},
year = {{2024}},
}