Characteristics of C-terminal, β-amyloid peptide binding fragment of neuroprotective protease inhibitor, cystatin C

Marta Spodzieja , Katarzyna Kalejta , Aleksandra Kołodziejczyk , Martyna Maszota-Zieleniak , Sylwia Rodziewicz-Motowidło , Wioletta Żmudzińska , Paulina Czaplewska


Cystatin C originally identified as a cysteine proteases inhibitor has a broad spectrum of biolog- ical roles ranging from inhibition of extracellular cysteine protease activities, bone resorption, and modulation of inflammatory responses to stimulation of fibroblasts proliferation. There is an increasing number of evidence to suggest that human cystatin C (hCC) might play a protective role in the pathophysiology of sporadic Alzheimer ’ s disease. In vivo and in vitro results well documented the association of hCC with A β and the hCC ‐ induced inhibition of A β fibril formation. In our earlier work, using a combination of selective proteolytic methods and MS spectroscopy, C ‐ terminal fragment hCC(101 ‐ 117) was identified as the A β ‐ binding region. The fragment of A β peptide responsible for the complex formation with hCC was found in the middle, highly hydrophobic part, A β (17 ‐ 24). Structures and affinities of both A β and hCC binding sites were characterized by the enzyme ‐ linked immunosorbent assay ‐ like assay, by surface plasmon resonance, and by nano ‐ ESI ‐ FTICR MS of the hCC – A β ‐ binding peptide complexes. In the in vitro inhibition studies, the binding cystatin sequence, hCC(101 ‐ 117), revealed the highest relative inhibitory effect toward A β ‐ fibril formation. Herein, we present further studies on molecular details of the hCC ‐ A β complex. With Ala substi- tution, affinity experiments, and enzyme ‐ linked immunosorbent assay ‐ like assays for the A β ‐ binding fragment, hCC(101 ‐ 117), and its variants, the importance of individual amino acid resi- dues for the protein interaction was evaluated. The results were analyzed using hCC(101 ‐ 117) nuclear magnetic resonance structural data with molecular dynamics calculations and molecular modeling of the complexes. The results point to conformational requirements and special impor- tance of some amino acid residues for the protein interaction. The obtained results might be helpful for the design of low molecular compounds modulating the biological role of both proteins. Copyright © 2016 John Wiley & Sons, Ltd
Author Marta Spodzieja (FCh / DBCh / LMCh)
Marta Spodzieja,,
- Laboratory of Medical Chemistry
, Katarzyna Kalejta (FCh / DBCh / LMCh)
Katarzyna Kalejta,,
- Laboratory of Medical Chemistry
, Aleksandra Kołodziejczyk (FCh / DBCh / LMCh)
Aleksandra Kołodziejczyk,,
- Laboratory of Medical Chemistry
, Martyna Maszota-Zieleniak (FCh / DBCh / LMCh)
Martyna Maszota-Zieleniak,,
- Laboratory of Medical Chemistry
, Sylwia Rodziewicz-Motowidło (FCh / DBCh / LMCh)
Sylwia Rodziewicz-Motowidło,,
- Laboratory of Medical Chemistry
, Wioletta Żmudzińska (IFB / IB / LBiopolS)
Wioletta Żmudzińska,,
- Laboratory of Biopolymers Structure
, Paulina Czaplewska (IFB / M020 / DMCB)
Paulina Czaplewska,,
- Department of Molecular and Cellular Biology
Journal seriesJournal of Molecular Recognition, ISSN 0952-3499, (A 25 pkt)
Issue year2017
Publication size in sheets0.6
Article numbere2581
Keywords in EnglishAlzheimer' s disease, amyloid, Aβ, cerebral amyloidosis, cystatin C, neurodegeneration
ASJC Classification1312 Molecular Biology; 1315 Structural Biology
Languageen angielski
Not used for evaluationyes
Score (nominal)0
Publication indicators WoS Citations = 2; Scopus Citations = 3; Scopus SNIP (Source Normalised Impact per Paper): 2017 = 0.686; WoS Impact Factor: 2017 = 1.868 (2) - 2017=1.901 (5)
Citation count*4 (2020-05-29)
Additional fields
UwagiArtykuł został zgłoszony do parametryzacji za lata 2013-2016
Share Share

Get link to the record

* presented citation count is obtained through Internet information analysis and it is close to the number calculated by the Publish or Perish system.
Are you sure?