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Crystal structure of human cystatin C stabilized against amyloid formation

Kolodziejczyk, Robert; Michalska, Karolina; Hernandez-Santoyo, Alejandra; Wahlbom, Maria LU ; Grubb, Anders LU and Jaskolski, Mariusz (2010) In The FEBS Journal 277(7). p.1726-1737
Abstract
Human cystatin C (HCC) is a family 2 cystatin inhibitor of papain-like (C1) and legumain-related (C13) cysteine proteases. In pathophysiological processes, the nature of which is not understood, HCC is codeposited in the amyloid plaques of Alzheimer's disease or Down's syndrome. The amyloidogenic properties of HCC are greatly increased in a naturally occurring L68Q variant, resulting in fatal cerebral amyloid angiopathy in early adult life. In all crystal structures of cystatin C studied to date, the protein has been found to form 3D domain-swapped dimers, created through a conformational change of a beta-hairpin loop, L1, from the papain-binding epitope. We have created monomer-stabilized human cystatin C, with an engineered disulfide... (More)
Human cystatin C (HCC) is a family 2 cystatin inhibitor of papain-like (C1) and legumain-related (C13) cysteine proteases. In pathophysiological processes, the nature of which is not understood, HCC is codeposited in the amyloid plaques of Alzheimer's disease or Down's syndrome. The amyloidogenic properties of HCC are greatly increased in a naturally occurring L68Q variant, resulting in fatal cerebral amyloid angiopathy in early adult life. In all crystal structures of cystatin C studied to date, the protein has been found to form 3D domain-swapped dimers, created through a conformational change of a beta-hairpin loop, L1, from the papain-binding epitope. We have created monomer-stabilized human cystatin C, with an engineered disulfide bond (L47C)-(G69C) between the structural elements that become separated upon domain swapping. The mutant has drastically reduced dimerization and fibril formation properties, but its inhibition of papain is unaltered. The structure confirms the success of the protein engineering experiment to abolish 3D domain swapping and, in consequence, amyloid fibril formation. It illustrates for the first time the fold of monomeric cystatin C and allows verification of earlier predictions based on the domain-swapped forms and on the structure of chicken cystatin. Importantly, the structure defines the so-far unknown conformation of loop L1, which is essential for the inhibition of papain-like cysteine proteases. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
engineering, protein, cysteine protease inhibitor, amyloid, crystal twinning, 3D domain swapping
in
The FEBS Journal
volume
277
issue
7
pages
1726 - 1737
publisher
Federation of European Neuroscience Societies and Blackwell Publishing Ltd
external identifiers
  • wos:000275634500012
  • scopus:77949588305
ISSN
1742-464X
DOI
10.1111/j.1742-4658.2010.07596.x
language
English
LU publication?
yes
id
5f44ff0c-1b4b-4ee9-ad43-617fe93ec54e (old id 1588028)
date added to LUP
2010-04-22 11:08:07
date last changed
2018-05-29 12:05:15
@article{5f44ff0c-1b4b-4ee9-ad43-617fe93ec54e,
  abstract     = {Human cystatin C (HCC) is a family 2 cystatin inhibitor of papain-like (C1) and legumain-related (C13) cysteine proteases. In pathophysiological processes, the nature of which is not understood, HCC is codeposited in the amyloid plaques of Alzheimer's disease or Down's syndrome. The amyloidogenic properties of HCC are greatly increased in a naturally occurring L68Q variant, resulting in fatal cerebral amyloid angiopathy in early adult life. In all crystal structures of cystatin C studied to date, the protein has been found to form 3D domain-swapped dimers, created through a conformational change of a beta-hairpin loop, L1, from the papain-binding epitope. We have created monomer-stabilized human cystatin C, with an engineered disulfide bond (L47C)-(G69C) between the structural elements that become separated upon domain swapping. The mutant has drastically reduced dimerization and fibril formation properties, but its inhibition of papain is unaltered. The structure confirms the success of the protein engineering experiment to abolish 3D domain swapping and, in consequence, amyloid fibril formation. It illustrates for the first time the fold of monomeric cystatin C and allows verification of earlier predictions based on the domain-swapped forms and on the structure of chicken cystatin. Importantly, the structure defines the so-far unknown conformation of loop L1, which is essential for the inhibition of papain-like cysteine proteases.},
  author       = {Kolodziejczyk, Robert and Michalska, Karolina and Hernandez-Santoyo, Alejandra and Wahlbom, Maria and Grubb, Anders and Jaskolski, Mariusz},
  issn         = {1742-464X},
  keyword      = {engineering,protein,cysteine protease inhibitor,amyloid,crystal twinning,3D domain swapping},
  language     = {eng},
  number       = {7},
  pages        = {1726--1737},
  publisher    = {Federation of European Neuroscience Societies and Blackwell Publishing Ltd},
  series       = {The FEBS Journal},
  title        = {Crystal structure of human cystatin C stabilized against amyloid formation},
  url          = {http://dx.doi.org/10.1111/j.1742-4658.2010.07596.x},
  volume       = {277},
  year         = {2010},
}