Stabilization, Characterization and Selective Removal of Cystatin C Amyloid Oligomers.
Ranheimer Östner, Gustav ; Lindström, Veronica LU
; Hjort Christensen, Per
; Kozak, Maciej
; Abrahamson, Magnus
LU
and Grubb, Anders
LU
(2013)
In Journal of Biological Chemistry
288(23).
p.16438-16450
- Abstract
- The pathophysiological process in amyloid disorders usually involves the transformation of a functional monomeric protein via potentially toxic oligomers into amyloid fibrils. The structure and properties of the intermediary oligomers have been difficult to study due to their instability and dynamic equilibrium with smaller and larger species. In hereditary cystatin C amyloid angiopathy, a cystatin C variant is deposited in arterial walls and cause brain hemorrhage in young adults. In the present investigation, we use redox experiments of monomeric cystatin C, stabilized against domain swapping by an intramolecular disulfide bond, to generate stable oligomers (dimers, trimers, tetramers, decamers and high molecular weight oligomers). These... (More)
- The pathophysiological process in amyloid disorders usually involves the transformation of a functional monomeric protein via potentially toxic oligomers into amyloid fibrils. The structure and properties of the intermediary oligomers have been difficult to study due to their instability and dynamic equilibrium with smaller and larger species. In hereditary cystatin C amyloid angiopathy, a cystatin C variant is deposited in arterial walls and cause brain hemorrhage in young adults. In the present investigation, we use redox experiments of monomeric cystatin C, stabilized against domain swapping by an intramolecular disulfide bond, to generate stable oligomers (dimers, trimers, tetramers, decamers and high molecular weight oligomers). These oligomers were characterized concerning size by gel filtration, polyacrylamide gel electrophoresis and mass spectrometry, concerning shape by electron and atomic force microscopy, and, concerning function, by assays of their capacity to inhibit proteases. The results showed the oligomers to be highly ordered, domainswapped assemblies of cystatin C and that the oligomers could not build larger oligomers, or fibrils, without domain swapping. The stabilized oligomers were used to induce antibody formation in rabbits. After immunosorption, using immobilized monomeric cystatin C, and elution from columns with immobilized cystatin C oligomers, oligomer-specific antibodies were obtained. These could be used to selectively remove cystatin C dimers from biological fluids containing both dimers and monomers. (Less)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/3805214
- author
- Ranheimer Östner, Gustav
; Lindström, Veronica
LU
; Hjort Christensen, Per
; Kozak, Maciej
; Abrahamson, Magnus
LU
and Grubb, Anders
LU
- organization
- publishing date
- 2013
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 288
- issue
- 23
- pages
- 16438 - 16450
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- wos:000320378900021
- pmid:23629649
- scopus:84878769258
- pmid:23629649
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M113.469593
- language
- English
- LU publication?
- yes
- id
- 01485e37-9a79-4087-8ef1-5d1c9162e641 (old id 3805214)
- alternative location
- http://www.ncbi.nlm.nih.gov/pubmed/23629649?dopt=Abstract
- date added to LUP
- 2016-04-01 10:42:16
- date last changed
- 2023-01-02 06:58:14
@article{01485e37-9a79-4087-8ef1-5d1c9162e641,
abstract = {{The pathophysiological process in amyloid disorders usually involves the transformation of a functional monomeric protein via potentially toxic oligomers into amyloid fibrils. The structure and properties of the intermediary oligomers have been difficult to study due to their instability and dynamic equilibrium with smaller and larger species. In hereditary cystatin C amyloid angiopathy, a cystatin C variant is deposited in arterial walls and cause brain hemorrhage in young adults. In the present investigation, we use redox experiments of monomeric cystatin C, stabilized against domain swapping by an intramolecular disulfide bond, to generate stable oligomers (dimers, trimers, tetramers, decamers and high molecular weight oligomers). These oligomers were characterized concerning size by gel filtration, polyacrylamide gel electrophoresis and mass spectrometry, concerning shape by electron and atomic force microscopy, and, concerning function, by assays of their capacity to inhibit proteases. The results showed the oligomers to be highly ordered, domainswapped assemblies of cystatin C and that the oligomers could not build larger oligomers, or fibrils, without domain swapping. The stabilized oligomers were used to induce antibody formation in rabbits. After immunosorption, using immobilized monomeric cystatin C, and elution from columns with immobilized cystatin C oligomers, oligomer-specific antibodies were obtained. These could be used to selectively remove cystatin C dimers from biological fluids containing both dimers and monomers.}},
author = {{Ranheimer Östner, Gustav and Lindström, Veronica and Hjort Christensen, Per and Kozak, Maciej and Abrahamson, Magnus and Grubb, Anders}},
issn = {{1083-351X}},
language = {{eng}},
number = {{23}},
pages = {{16438--16450}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Stabilization, Characterization and Selective Removal of Cystatin C Amyloid Oligomers.}},
url = {{https://lup.lub.lu.se/search/files/2067698/4219788.pdf}},
doi = {{10.1074/jbc.M113.469593}},
volume = {{288}},
year = {{2013}},
}