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

Magdalena Murawska , Aneta Szymańska , Anders Grubb , Maciej Kozak


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.
Author Magdalena Murawska - [Uniwersytet im. Adama Mickiewicza w Poznaniu]
Magdalena Murawska,,
- Uniwersytet im. Adama Mickiewicza w Poznaniu
, Aneta Szymańska (FCh / DBCh / LMCh)
Aneta Szymańska,,
- Laboratory of Medical Chemistry
, Anders Grubb - [Skånes universitetssjukhus]
Anders Grubb,,
, Maciej Kozak - [Uniwersytet im. Adama Mickiewicza w Poznaniu]
Maciej Kozak,,
- Uniwersytet im. Adama Mickiewicza w Poznaniu
Journal seriesNuclear Instruments & Methods in Physics Research. Section B: Beam Interactions With Materials and Atoms, ISSN 0168-583X, (A 25 pkt)
Issue year2017
Publication size in sheets0.5
Keywords in Englishhuman cystatin C, shape determination, small angle X-ray scattering, synchrotron radiation
ASJC Classification3105 Instrumentation; 3106 Nuclear and High Energy Physics
URL https://doi.org/10.1016/j.nimb.2017.02.090
Languageen angielski
Score (nominal)25
Score sourcejournalList
ScoreMinisterial score = 25.0, 29-05-2020, ArticleFromJournal
Publication indicators WoS Citations = 1; Scopus Citations = 1; Scopus SNIP (Source Normalised Impact per Paper): 2016 = 0.906; WoS Impact Factor: 2017 = 1.323 (2) - 2017=1.297 (5)
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