Advanced

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)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Journal of Biological Chemistry
volume
279
issue
23
pages
10 pages
publisher
ASBMB
external identifiers
  • wos:000221702500048
  • scopus:2642567686
ISSN
1083-351X
DOI
10.1074/jbc.M402621200
language
English
LU publication?
yes
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
2007-07-05 15:07:19
date last changed
2017-11-19 03:31:57
@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    = {ASBMB},
  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          = {http://dx.doi.org/10.1074/jbc.M402621200},
  volume       = {279},
  year         = {2004},
}