NMR and crystallographic structural studies of the extremely stable monomeric variant of human cystatin C with single amino acid substitution

Martyna Maszota-Zieleniak , Przemysław Jurczak , Marta Orlikowska , Igor Zhukov , Dominika Borek , Zbyszek Otwinowski , Piotr Skowron , Zuzanna Pietralik , Maciej Kozak , Aneta Szymańska , Sylwia Rodziewicz-Motowidło

Abstract

Human cystatin C (hCC), a member of the superfamily of papain-like cysteine protease inhibitors, is the most widespread cystatin in human body fluids. This small protein, in addition to its physiological function, is involved in various diseases, including cerebral amyloid angiopathy, cerebral hemorrhage, stroke, and dementia. Physiologically active hCC is a monomer. However, all structural studies based on crystallization led to the dimeric structure formed as a result of a three-dimensional exchange of the protein domains (3D domain swapping). The monomeric structure was obtained only for hCC variant V57N and for the protein stabilized by an additional disulfide bridge. With this study, we extend the number of models of monomeric hCC by an additional hCC variant with a single amino acid substitution in the flexible loop L1. The V57G variant was chosen for the X-ray and NMR structural analysis due to its exceptional conformational stability in solution. In this work, we show for the first time the structural and dynamics studies of human cystatin C variant in solution. We were also able to compare these data with the crystal structure of the hCC V57G and with other cystatins. The overall cystatin fold is retained in the solute form. Additionally, structural information concerning the N terminus was obtained during our studies and presented for the first time.
Publication typeIn press (online first, early view)
Author Martyna Maszota-Zieleniak (FCh / DBCh / LMCh)
Martyna Maszota-Zieleniak,,
- Laboratory of Medical Chemistry
, Przemysław Jurczak (FCh / DBCh / LMCh)
Przemysław Jurczak,,
- Laboratory of Medical Chemistry
, Marta Orlikowska (FCh / DBCh / LMCh)
Marta Orlikowska,,
- Laboratory of Medical Chemistry
, Igor Zhukov - [Uniwersytet im. Adama Mickiewicza w Poznaniu]
Igor Zhukov,,
-
- Uniwersytet im. Adama Mickiewicza w Poznaniu
, Dominika Borek - [UT Southwestern Medical School]
Dominika Borek,,
-
-
, Zbyszek Otwinowski - [UT Southwestern Medical School]
Zbyszek Otwinowski,,
-
-
, Piotr Skowron (FCh / DMBt / LGE)
Piotr Skowron,,
- Laboratory of Genetic Engineering
, Zuzanna Pietralik - [Uniwersytet im. Adama Mickiewicza w Poznaniu]
Zuzanna Pietralik,,
-
- Uniwersytet im. Adama Mickiewicza w Poznaniu
, Maciej Kozak - [Uniwersytet im. Adama Mickiewicza w Poznaniu]
Maciej Kozak,,
-
- Uniwersytet im. Adama Mickiewicza w Poznaniu
, Aneta Szymańska (FCh / DBCh / LMCh)
Aneta Szymańska,,
- Laboratory of Medical Chemistry
et al.`
Journal seriesFEBS Journal, ISSN 1742-464X, (N/A 100 pkt)
Issue year2019
Noonline first
Pages1-15
Publication size in sheets0.7
Keywords in EnglishhCC V57G variant, human cystatin C, NMR, protein structure, X-ray
ASJC Classification1307 Cell Biology; 1312 Molecular Biology; 1303 Biochemistry
DOIDOI:10.1111/febs.15010
URL https://doi.org/10.1111/febs.15010
Languageen angielski
Score (nominal)100
Score sourcejournalList
ScoreMinisterial score = 100.0, 02-12-2019, ArticleFromJournal
Publication indicators Scopus Citations = 0; WoS Citations = 0; Scopus SNIP (Source Normalised Impact per Paper): 2018 = 1.148; WoS Impact Factor: 2018 = 4.739 (2) - 2018=4.432 (5)
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