A label-free high-throughput protein solubility assay and its application to Aβ40
(2024) In Biophysical Chemistry- Abstract
- A major hallmark of Alzheimer's disease is the accumulation of aggregated amyloid peptide (Aβ) in the brain. Here we develop a solubility assay for proteins and measure the solubility of Aβ40. In brief, the method utilizes 96-well filter plates to separate monomeric Aβ from aggregated Aβ, and the small species are quantified with the amine reactive dye o-phthalaldehyde (OPA). This procedure ensures that solubility is measured for unlabeled species, and makes the assay high-throughput and inexpensive. We demonstrate that the filter plates successfully separate fibrils from monomer, with negligible monomer adsorption, and that OPA can quantify Aβ peptides in a concentration range from 44 nM to 20 μM. We also show that adding a methionine... (More)
- A major hallmark of Alzheimer's disease is the accumulation of aggregated amyloid peptide (Aβ) in the brain. Here we develop a solubility assay for proteins and measure the solubility of Aβ40. In brief, the method utilizes 96-well filter plates to separate monomeric Aβ from aggregated Aβ, and the small species are quantified with the amine reactive dye o-phthalaldehyde (OPA). This procedure ensures that solubility is measured for unlabeled species, and makes the assay high-throughput and inexpensive. We demonstrate that the filter plates successfully separate fibrils from monomer, with negligible monomer adsorption, and that OPA can quantify Aβ peptides in a concentration range from 44 nM to 20 μM. We also show that adding a methionine residue to the N-terminus of Aβ1–40 decreases the solubility by <3-fold. The method will facilitate further solubility studies, and contribute to the understanding of the thermodynamics of amyloid fibril formation. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/59d15558-9c4d-4120-a07f-e0d52403ef99
- author
- Lindberg, Max LU ; Axell, Emil LU ; Sparr, Emma LU and Snogerup-Linse, Sara LU
- organization
- publishing date
- 2024-01-02
- type
- Contribution to journal
- publication status
- epub
- subject
- in
- Biophysical Chemistry
- publisher
- Elsevier
- external identifiers
-
- scopus:85184008680
- ISSN
- 0301-4622
- DOI
- 10.1016/j.bpc.2023.107165
- language
- English
- LU publication?
- yes
- id
- 59d15558-9c4d-4120-a07f-e0d52403ef99
- date added to LUP
- 2024-01-29 10:22:29
- date last changed
- 2024-03-05 12:12:54
@article{59d15558-9c4d-4120-a07f-e0d52403ef99, abstract = {{A major hallmark of Alzheimer's disease is the accumulation of aggregated amyloid peptide (Aβ) in the brain. Here we develop a solubility assay for proteins and measure the solubility of Aβ40. In brief, the method utilizes 96-well filter plates to separate monomeric Aβ from aggregated Aβ, and the small species are quantified with the amine reactive dye o-phthalaldehyde (OPA). This procedure ensures that solubility is measured for unlabeled species, and makes the assay high-throughput and inexpensive. We demonstrate that the filter plates successfully separate fibrils from monomer, with negligible monomer adsorption, and that OPA can quantify Aβ peptides in a concentration range from 44 nM to 20 μM. We also show that adding a methionine residue to the N-terminus of Aβ1–40 decreases the solubility by <3-fold. The method will facilitate further solubility studies, and contribute to the understanding of the thermodynamics of amyloid fibril formation.}}, author = {{Lindberg, Max and Axell, Emil and Sparr, Emma and Snogerup-Linse, Sara}}, issn = {{0301-4622}}, language = {{eng}}, month = {{01}}, publisher = {{Elsevier}}, series = {{Biophysical Chemistry}}, title = {{A label-free high-throughput protein solubility assay and its application to A<i>β</i>40}}, url = {{http://dx.doi.org/10.1016/j.bpc.2023.107165}}, doi = {{10.1016/j.bpc.2023.107165}}, year = {{2024}}, }