Advanced

Structural characterization of covalently stabilized human cystatin c oligomers

Chrabaszczewska, Magdalena ; Sieradzan, Adam K. ; Rodziewicz-Motowidło, Sylwia ; Grubb, Anders LU ; Dobson, Christopher M. ; Kumita, Janet R. and Kozak, Maciej (2020) In International Journal of Molecular Sciences 21(16).
Abstract

Human cystatin C (HCC), a cysteine-protease inhibitor, exists as a folded monomer under physiological conditions but has the ability to self-assemble via domain swapping into multimeric states, including oligomers with a doughnut-like structure. The structure of the monomeric HCC has been solved by X-ray crystallography, and a covalently linked version of HCC (stab-1 HCC) is able to form stable oligomeric species containing 10–12 monomeric subunits. We have performed molecular modeling, and in conjunction with experimental parameters obtained from atomic force microscopy (AFM), transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS) measurements, we observe that the structures are essentially flat, with a height... (More)

Human cystatin C (HCC), a cysteine-protease inhibitor, exists as a folded monomer under physiological conditions but has the ability to self-assemble via domain swapping into multimeric states, including oligomers with a doughnut-like structure. The structure of the monomeric HCC has been solved by X-ray crystallography, and a covalently linked version of HCC (stab-1 HCC) is able to form stable oligomeric species containing 10–12 monomeric subunits. We have performed molecular modeling, and in conjunction with experimental parameters obtained from atomic force microscopy (AFM), transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS) measurements, we observe that the structures are essentially flat, with a height of about 2 nm, and the distance between the outer edge of the ring and the edge of the central cavity is ~5.1 nm. These dimensions correspond to the height and diameter of one stab-1 HCC subunit and we present a dodecamer model for stabilized cystatin C oligomers using molecular dynamics simulations and experimentally measured parameters. Given that oligomeric species in protein aggregation reactions are often transient and very highly heterogeneous, the structural information presented here on these isolated stab-1 HCC oligomers may be useful to further explore the physiological relevance of different structural species of cystatin C in relation to protein misfolding disease.

(Less)
Please use this url to cite or link to this publication:
author
; ; ; ; ; and
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Cystatin C, Domain swapping, Oligomers, Protein misfolding
in
International Journal of Molecular Sciences
volume
21
issue
16
article number
5860
pages
17 pages
publisher
MDPI AG
external identifiers
  • pmid:32824145
  • scopus:85089603448
ISSN
1661-6596
DOI
10.3390/ijms21165860
language
English
LU publication?
yes
id
fa057986-b016-4ddb-b7f9-3ed6238128fa
date added to LUP
2020-08-27 13:25:20
date last changed
2021-02-17 04:49:56
@article{fa057986-b016-4ddb-b7f9-3ed6238128fa,
  abstract     = {<p>Human cystatin C (HCC), a cysteine-protease inhibitor, exists as a folded monomer under physiological conditions but has the ability to self-assemble via domain swapping into multimeric states, including oligomers with a doughnut-like structure. The structure of the monomeric HCC has been solved by X-ray crystallography, and a covalently linked version of HCC (stab-1 HCC) is able to form stable oligomeric species containing 10–12 monomeric subunits. We have performed molecular modeling, and in conjunction with experimental parameters obtained from atomic force microscopy (AFM), transmission electron microscopy (TEM) and small-angle X-ray scattering (SAXS) measurements, we observe that the structures are essentially flat, with a height of about 2 nm, and the distance between the outer edge of the ring and the edge of the central cavity is ~5.1 nm. These dimensions correspond to the height and diameter of one stab-1 HCC subunit and we present a dodecamer model for stabilized cystatin C oligomers using molecular dynamics simulations and experimentally measured parameters. Given that oligomeric species in protein aggregation reactions are often transient and very highly heterogeneous, the structural information presented here on these isolated stab-1 HCC oligomers may be useful to further explore the physiological relevance of different structural species of cystatin C in relation to protein misfolding disease.</p>},
  author       = {Chrabaszczewska, Magdalena and Sieradzan, Adam K. and Rodziewicz-Motowidło, Sylwia and Grubb, Anders and Dobson, Christopher M. and Kumita, Janet R. and Kozak, Maciej},
  issn         = {1661-6596},
  language     = {eng},
  number       = {16},
  publisher    = {MDPI AG},
  series       = {International Journal of Molecular Sciences},
  title        = {Structural characterization of covalently stabilized human cystatin c oligomers},
  url          = {http://dx.doi.org/10.3390/ijms21165860},
  doi          = {10.3390/ijms21165860},
  volume       = {21},
  year         = {2020},
}