Chronopotentiometric sensing of anterior gradient 2 protein

0476126 - BFÚ 2018 RIV GB eng J - Článek v odborném periodiku
Vargová, Veronika - Hrstka, R. - Hernychová, L. - Coufalová, D. - Ostatná, Veronika
Chronopotentiometric sensing of anterior gradient 2 protein.
Electrochimica acta. Roč. 240, JUN2017 (2017), s. 250-257. ISSN 0013-4686. E-ISSN 1873-3859
Grant CEP: GA ČR(CZ) GA13-00956S
Institucionální podpora: RVO:68081707
Klíčová slova: catalyzed hydrogen evolution * mercury-electrodes * affinity tags
Obor OECD: Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)
Impakt faktor: 5.116, rok: 2017

The study of new proteins and particularly those involved in cancer development is still a focal point of interest. We studied the anterior gradient 2 (AGR2) oncoprotein, aberrantly expressed in a number of human cancers. For the first time the electrochemical behaviour of various variants of the AGR2 was described using constant current chronopotentiometric stripping (CPS) analysis at mercury electrodes. In the first part, we show that mutation of AGR2 protein at its sole cysteine significantly changed its CPS response compared to wild type AGR2, probably due to their different adsorption and some deviations in their structures, which were obtained by analysis of hydrogen deuterium (HID) exchange connected with high resolution mass spectrometry (MS). In the second part we studied the influence of His-tag modification, widely used in the purification of recombinant proteins, on CPS response and H/D exchange. Addition of a His-tag, containing positively charged and electroactive residues, affected the CPS peak H of His(6)-tagged AGR2 compared to non-tagged AGR2 protein due to different adsorption as well as variation in the structure at the negatively charged interface. H/D exchange MS analysis confirmed differences in the structure of these two variants even in solution. The uncovering of AGR2 behaviour at charged surfaces gives us the opportunity to study its interactions with other biomedically important proteins. (C) 2017 Elsevier Ltd. All rights reserved.
Trvalý link: http://hdl.handle.net/11104/0272678