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Overall conformation of covalently stabilized domain-swapped dimer of human cystatin C in solution

Murawska, Magdalena; Szymańska, Aneta; Grubb, Anders LU and Kozak, Maciej (2017) In Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms 411. p.136-140
Abstract

Human cystatin C (HCC), a small protein, plays a crucial role in inhibition of cysteine proteases. The most common structural form of human cystatin C in crystals is a dimer, which has been evidenced both for the native protein and its mutants. In these structures, HCC dimers were formed through the mechanism of domain swapping. The structure of the monomeric form of human cystatin C was determined for V57N mutant and the mutant with the engineered disulfide bond (L47C)–(G69C) (known as stab1-HCC). On the basis of stab1-HCC, a number of covalently stabilized oligomers, including also dimers have been obtained. The aim of this study was to analyze the structure of the covalently stabilized dimer HCC in solution by the small angle X-ray... (More)

Human cystatin C (HCC), a small protein, plays a crucial role in inhibition of cysteine proteases. The most common structural form of human cystatin C in crystals is a dimer, which has been evidenced both for the native protein and its mutants. In these structures, HCC dimers were formed through the mechanism of domain swapping. The structure of the monomeric form of human cystatin C was determined for V57N mutant and the mutant with the engineered disulfide bond (L47C)–(G69C) (known as stab1-HCC). On the basis of stab1-HCC, a number of covalently stabilized oligomers, including also dimers have been obtained. The aim of this study was to analyze the structure of the covalently stabilized dimer HCC in solution by the small angle X-ray scattering (SAXS) technique and synchrotron radiation. Experimental data confirmed that in solution this protein forms a dimer, which is characterized by the radius of gyration RG = 3.1 nm and maximum intramolecular distance Dmax = 10.3 nm. Using the ab initio method and program DAMMIN, we propose a low resolution structure of stabilized covalently cystatin C in solution. Stab-HCC dimer adopts in solution an elongated conformation, which is well reconstructed by the ab initio model.

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publishing date
type
Contribution to journal
publication status
published
subject
keywords
Human cystatin C, Shape determination, Small angle X-ray scattering, Synchrotron radiation
in
Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms
volume
411
pages
5 pages
publisher
Elsevier
external identifiers
  • scopus:85014678969
ISSN
0168-583X
DOI
10.1016/j.nimb.2017.02.090
language
English
LU publication?
no
id
b646da39-771c-4105-b100-dec12cc3d6d2
date added to LUP
2017-11-22 08:47:45
date last changed
2018-05-27 04:47:33
@article{b646da39-771c-4105-b100-dec12cc3d6d2,
  abstract     = {<p>Human cystatin C (HCC), a small protein, plays a crucial role in inhibition of cysteine proteases. The most common structural form of human cystatin C in crystals is a dimer, which has been evidenced both for the native protein and its mutants. In these structures, HCC dimers were formed through the mechanism of domain swapping. The structure of the monomeric form of human cystatin C was determined for V57N mutant and the mutant with the engineered disulfide bond (L47C)–(G69C) (known as stab1-HCC). On the basis of stab1-HCC, a number of covalently stabilized oligomers, including also dimers have been obtained. The aim of this study was to analyze the structure of the covalently stabilized dimer HCC in solution by the small angle X-ray scattering (SAXS) technique and synchrotron radiation. Experimental data confirmed that in solution this protein forms a dimer, which is characterized by the radius of gyration R<sub>G</sub> = 3.1 nm and maximum intramolecular distance D<sub>max</sub> = 10.3 nm. Using the ab initio method and program DAMMIN, we propose a low resolution structure of stabilized covalently cystatin C in solution. Stab-HCC dimer adopts in solution an elongated conformation, which is well reconstructed by the ab initio model.</p>},
  author       = {Murawska, Magdalena and Szymańska, Aneta and Grubb, Anders and Kozak, Maciej},
  issn         = {0168-583X},
  keyword      = {Human cystatin C,Shape determination,Small angle X-ray scattering,Synchrotron radiation},
  language     = {eng},
  month        = {11},
  pages        = {136--140},
  publisher    = {Elsevier},
  series       = {Nuclear Instruments and Methods in Physics Research, Section B: Beam Interactions with Materials and Atoms},
  title        = {Overall conformation of covalently stabilized domain-swapped dimer of human cystatin C in solution},
  url          = {http://dx.doi.org/10.1016/j.nimb.2017.02.090},
  volume       = {411},
  year         = {2017},
}