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
AbstractHuman 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.
|Journal series||Amino Acids, ISSN 0939-4451, (A 25 pkt)|
|Publication size in sheets||0.55|
|Keywords in English||human cystatin C, HDX exchange, mass spectrometry, epitope identification|
|License||Other; published final; ; with publication|
|Score|| = 25.0, 20-12-2017, ArticleFromJournal|
= 30.0, 20-12-2017, ArticleFromJournal
|Publication indicators||: 2016 = 3.173 (2) - 2016=3.241 (5)|
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