Number of the records: 1

Electrochemical analysis of Os(VI)-modified glycoproteins and label-free glycoprotein detection eluted from lectin capillary column

SYSNO ASEP	0476223
Document Type	J - Journal Article
R&D Document Type	Journal Article
Subsidiary J	Článek ve WOS
Title	Electrochemical analysis of Os(VI)-modified glycoproteins and label-free glycoprotein detection eluted from lectin capillary column
Author(s)	Trefulka, Mojmír (BFU-R)_{RID, ORCID} Dorčák, Vlastimil (BFU-R)_{RID, ORCID} Křenková, Jana (UIACH-O)_{RID, ORCID} Foret, František (UIACH-O)_{RID, ORCID} Paleček, Emil (BFU-R)_{RID, ORCID}
Number of authors	5
Source Title	Electrochimica acta. - : Elsevier - ISSN 0013-4686 Roč. 239, JUN 2017 (2017), s. 10-15
Number of pages	6 s.
Publication form	Print - P
Language	eng - English
Country	GB - United Kingdom
Keywords	voltammetric determination ; modified electrodes ; high-sensitivity ; concanavalin-a
Subject RIV	CG - Electrochemistry
OECD category	Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)
Subject RIV - cooperation	Institute of Analytical Chemistry - Analytical Chemistry, Separation
R&D Projects	GA15-15479S GA ČR - Czech Science Foundation (CSF)
Institutional support	BFU-R - RVO:68081707 ; UIACH-O - RVO:68081715
UT WOS	000401114000002
DOI	10.1016/j.electacta.2017.04.045
Annotation	Earlier we showed that using Os(VI) temed complex, glycans can be modified directly in glycoproteins and detected voltammetrically at mu M concentrations at carbon electrodes. Here we used Os(VI) 2,2'-bipyridine for modification of ribonucleases and Hg electrodes for voltammetric detection. We show that glycosylated RNase B produced electrocatalytic signal (after separation from the reaction mixture) at pM concentrations while non-glycosylated RNase A yielded negligible signal under the same conditions. Using Os(VI) temed and carbon electrodes voltammetric detection was less sensitive but allowed detection of RNase B in an excess of non-glycosylated protein, directly in the reaction mixture. We also showed that the constant current chronopotentiometric stripping (CPS) peak H (which is due to the ability of some amino acid residues in proteins to catalyze hydrogen evolution at Hg electrodes) could be used for protein structure-sensitive analysis at mercury electrodes. Using this peak, here we show that glycosylation of RNase stabilizes its molecule at the electrode. On the other hand, Os(VI) temed modification results in destabilization of this surface-attached protein. Peak H was also used for detection of RNase B separated from a large excess of non-glycosylated proteins on a lectin (concanavalin A) monolithic flow-through column. (C) 2017 Elsevier Ltd. All rights reserved.
Workplace	Institute of Biophysics
Contact	Jana Poláková, polakova@ibp.cz, Tel.: 541 517 244
Year of Publishing	2018

Number of the records: 1