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)
Please use this url to cite or link to this publication:
https://lup.lub.lu.se/record/1588028
- author
- Kolodziejczyk, Robert
; Michalska, Karolina
; Hernandez-Santoyo, Alejandra
; Wahlbom, Maria
LU
; Grubb, Anders
LU
and Jaskolski, Mariusz
- organization
- publishing date
- 2010
- 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
- Wiley-Blackwell
- external identifiers
-
- wos:000275634500012
- scopus:77949588305
- pmid:20175878
- 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
- 2016-04-01 13:38:17
- date last changed
- 2023-01-04 00:02:21
@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}},
keywords = {{engineering; protein; cysteine protease inhibitor; amyloid; crystal twinning; 3D domain swapping}},
language = {{eng}},
number = {{7}},
pages = {{1726--1737}},
publisher = {{Wiley-Blackwell}},
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}},
doi = {{10.1111/j.1742-4658.2010.07596.x}},
volume = {{277}},
year = {{2010}},
}