Prevention of domain swapping inhibits dimerization and amyloid fibril formation of cystatin C : use of engineered disulfide bridges, antibodies, and carboxymethylpapain to stabilize the monomeric form of cystatin C
Wahlbom, Maria LU ; Wang, Xin LU ; Rodziewicz-Motowidlo, Sylwia ; Janowski, Robert ; Lindström, Veronica LU
; Önnerfjord, Patrik
LU
; Westermark, Gunilla
; Grzonka, Zbigniew
; Jaskolski, Mariusz
and Grubb, Anders
LU
(2004)
In Journal of Biological Chemistry
279(23).
p.24236-24245
- Abstract
Amyloidogenic proteins like cystatin C and prion proteins have been shown to form dimers by exchange of subdomains of the monomeric proteins. This process, called "three-dimensional domain swapping," has also been suggested to play a part in the generation of amyloid fibrils. One variant of cystatin C, L68Q cystatin C, is highly amyloidogenic, and persons carrying the corresponding gene suffer from massive cerebral amyloidosis leading to brain hemorrhage and death in early adult life. The present work describes the production of two variants of wild type and L68Q cystatin C with disulfide bridges at positions selected to inhibit domain swapping without affecting the biological function of the four cystatin C variants as cysteine... (More)
Amyloidogenic proteins like cystatin C and prion proteins have been shown to form dimers by exchange of subdomains of the monomeric proteins. This process, called "three-dimensional domain swapping," has also been suggested to play a part in the generation of amyloid fibrils. One variant of cystatin C, L68Q cystatin C, is highly amyloidogenic, and persons carrying the corresponding gene suffer from massive cerebral amyloidosis leading to brain hemorrhage and death in early adult life. The present work describes the production of two variants of wild type and L68Q cystatin C with disulfide bridges at positions selected to inhibit domain swapping without affecting the biological function of the four cystatin C variants as cysteine protease inhibitors. The capacity of the four variant proteins to form dimers was tested and compared with that of wild type and L68Q cystatin C. In contrast to the latter two proteins, all four protein variants stabilized by disulfide bridges were resistant toward the formation of dimers. The capacity of the two stabilized variants of wild type cystatin C to form amyloid fibrils was investigated and found to be reduced by 80% compared with that of wild type cystatin C. In an effort to investigate whether exogenous agents could also suppress the formation of dimers of wild type and L68Q cystatin C, a monoclonal antibody or carboxymethylpapain, an inactivated form of a cysteine protease, was added to systems inducing dimerization of wild type and L68Q cystatin C. It was observed that catalytic amounts of both the monoclonal antibody and carboxymethylpapain could suppress dimerization.
(Less)
- author
- Wahlbom, Maria
LU
; Wang, Xin
LU
; Rodziewicz-Motowidlo, Sylwia
; Janowski, Robert
; Lindström, Veronica
LU
; Önnerfjord, Patrik
LU
; Westermark, Gunilla
; Grzonka, Zbigniew
; Jaskolski, Mariusz
and Grubb, Anders
LU
- organization
- publishing date
- 2004
- type
- Contribution to journal
- publication status
- published
- subject
- in
- Journal of Biological Chemistry
- volume
- 279
- issue
- 23
- pages
- 10 pages
- publisher
- American Society for Biochemistry and Molecular Biology
- external identifiers
-
- wos:000221702500048
- scopus:2642567686
- pmid:15028721
- ISSN
- 1083-351X
- DOI
- 10.1074/jbc.M402621200
- language
- English
- LU publication?
- yes
- additional info
- The information about affiliations in this record was updated in December 2015. The record was previously connected to the following departments: Cell and Matrix Biology (LUR000002), Division of Clinical Chemistry and Pharmacology (013250300), Connective Tissue Biology (013230151)
- id
- 64a410ed-cc4f-43f7-9161-8b31dcb36a7f (old id 121143)
- alternative location
- http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=15028721&dopt=Abstract
- date added to LUP
- 2016-04-01 12:02:26
- date last changed
- 2023-02-21 01:32:22
@article{64a410ed-cc4f-43f7-9161-8b31dcb36a7f,
abstract = {{<p>Amyloidogenic proteins like cystatin C and prion proteins have been shown to form dimers by exchange of subdomains of the monomeric proteins. This process, called "three-dimensional domain swapping," has also been suggested to play a part in the generation of amyloid fibrils. One variant of cystatin C, L68Q cystatin C, is highly amyloidogenic, and persons carrying the corresponding gene suffer from massive cerebral amyloidosis leading to brain hemorrhage and death in early adult life. The present work describes the production of two variants of wild type and L68Q cystatin C with disulfide bridges at positions selected to inhibit domain swapping without affecting the biological function of the four cystatin C variants as cysteine protease inhibitors. The capacity of the four variant proteins to form dimers was tested and compared with that of wild type and L68Q cystatin C. In contrast to the latter two proteins, all four protein variants stabilized by disulfide bridges were resistant toward the formation of dimers. The capacity of the two stabilized variants of wild type cystatin C to form amyloid fibrils was investigated and found to be reduced by 80% compared with that of wild type cystatin C. In an effort to investigate whether exogenous agents could also suppress the formation of dimers of wild type and L68Q cystatin C, a monoclonal antibody or carboxymethylpapain, an inactivated form of a cysteine protease, was added to systems inducing dimerization of wild type and L68Q cystatin C. It was observed that catalytic amounts of both the monoclonal antibody and carboxymethylpapain could suppress dimerization.</p>}},
author = {{Wahlbom, Maria and Wang, Xin and Rodziewicz-Motowidlo, Sylwia and Janowski, Robert and Lindström, Veronica and Önnerfjord, Patrik and Westermark, Gunilla and Grzonka, Zbigniew and Jaskolski, Mariusz and Grubb, Anders}},
issn = {{1083-351X}},
language = {{eng}},
number = {{23}},
pages = {{24236--24245}},
publisher = {{American Society for Biochemistry and Molecular Biology}},
series = {{Journal of Biological Chemistry}},
title = {{Prevention of domain swapping inhibits dimerization and amyloid fibril formation of cystatin C : use of engineered disulfide bridges, antibodies, and carboxymethylpapain to stabilize the monomeric form of cystatin C}},
url = {{https://lup.lub.lu.se/search/files/2754941/602579.pdf}},
doi = {{10.1074/jbc.M402621200}},
volume = {{279}},
year = {{2004}},
}