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

Abstract

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 PChM
Marta Spodzieja,,
- Laboratory of Medical Chemistry
, Katarzyna Kalejta PChM
Katarzyna Kalejta,,
- Laboratory of Medical Chemistry
, Aleksandra Kołodziejczyk PChM
Aleksandra Kołodziejczyk,,
- Laboratory of Medical Chemistry
, Martyna Maszota-Zieleniak PChM
Martyna Maszota-Zieleniak,,
- Laboratory of Medical Chemistry
, Sylwia Rodziewicz-Motowidło PChM
Sylwia Rodziewicz-Motowidło,,
- Laboratory of Medical Chemistry
, Wioletta Żmudzińska PSB
Wioletta Żmudzińska,,
- Laboratory of Biopolymers Structure
, Paulina Czaplewska KBMiK
Paulina Czaplewska,,
- Department of Molecular and Cellular Biology
Journal seriesJournal of Molecular Recognition, ISSN 0952-3499
Issue year2017
Vol30
No2
Pages1-13
Publication size in sheets0.6
Keywords in EnglishAlzheimer' s disease, amyloid, Aβ, cerebral amyloidosis, cystatin C, neurodegeneration
DOIDOI:10.1002/jmr.2581
URL http://onlinelibrary.wiley.com/doi/10.1002/jmr.2581/pdf
Languageen angielski
Score (nominal)25
ScoreMinisterial score = 25.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 25.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 2.175 (2) - 2016=2.051 (5)
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