Application of amide hydrogen/deuterium exchange mass spectrometry for epitope mapping in human cystatin C

Martyna Prądzińska , Izabela Behrendt , Juan Astorga-Wells , Aleksandr Manoilov , Roman A. Zubarev , Aleksandra Kołodziejczyk , Sylwia Rodziewicz-Motowidło , Paulina Czaplewska


Human cystatin C (hCC) is a small cysteine protease inhibitor whose oligomerization by propagated domain swapping is linked to certain neurological disorders. One of the ways to prevent hCC dimerization and fibrillogenesis is to enable its interaction with a proper antibody. Herein, the sites of interaction of hCC with dimer-preventing mouse monoclonal anti-hCC antibodies Cyst28 are studied and compared with the binding sites found for mAb Cyst10 that has almost no effect on hCC dimerization. In addition, hCC epitopes in complexes with native polyclonal antibodies extracted from human serum were studied. The results obtained with hydrogen–deuterium exchange mass spectrometry (HDX MS) were compared with the previous findings made using the excision/extraction MS approach. The main results from the two complementary MS-based approaches are found to be in agreement with each other, with some differences being attributed to the specificity of each method. The findings of the current studies may be important for future design of hCC dimerization inhibitors.
Author Martyna Prądzińska (FCh / DBCh / LMCh)
Martyna Prądzińska,,
- Laboratory of Medical Chemistry
, Izabela Behrendt (FCh / DBCh / LMCh)
Izabela Behrendt,,
- Laboratory of Medical Chemistry
, Juan Astorga-Wells
Juan Astorga-Wells,,
, Aleksandr Manoilov
Aleksandr Manoilov,,
, Roman A. Zubarev
Roman A. Zubarev,,
, Aleksandra Kołodziejczyk (FCh / DBCh / LMCh)
Aleksandra Kołodziejczyk,,
- Laboratory of Medical Chemistry
, Sylwia Rodziewicz-Motowidło (FCh / DBCh / LMCh)
Sylwia Rodziewicz-Motowidło,,
- Laboratory of Medical Chemistry
, Paulina Czaplewska (IFB / M020 / DMCB)
Paulina Czaplewska,,
- Department of Molecular and Cellular Biology
Journal seriesAmino Acids, ISSN 0939-4451, (A 25 pkt)
Issue year2016
Publication size in sheets0.55
Keywords in Englishhuman cystatin C, HDX exchange, mass spectrometry, epitope identification
Languageen angielski
LicenseOther; published final; Uznanie Autorstwa (CC-BY); with publication
Score (nominal)30
ScoreMinisterial score = 25.0, 20-12-2017, ArticleFromJournal
Ministerial score (2013-2016) = 30.0, 20-12-2017, ArticleFromJournal
Publication indicators WoS Impact Factor: 2016 = 3.173 (2) - 2016=3.241 (5)
Citation count*
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.