Počet záznamů: 1
Electrochemical analysis of Os(VI)-modified glycoproteins and label-free glycoprotein detection eluted from lectin capillary column
- 1.0476223 - BFU-R 2018 RIV GB eng J - Článek v odborném periodiku
Trefulka, Mojmír - Dorčák, Vlastimil - Křenková, Jana - Foret, František - Paleček, Emil
Electrochemical analysis of Os(VI)-modified glycoproteins and label-free glycoprotein detection eluted from lectin capillary column.
Electrochimica acta. Roč. 239, JUN 2017 (2017), s. 10-15. ISSN 0013-4686
Grant CEP: GA ČR(CZ) GA15-15479S
Institucionální podpora: RVO:68081707 ; RVO:68081715
Klíčová slova: voltammetric determination * modified electrodes * high-sensitivity * concanavalin-a
Kód oboru RIV: CG - Elektrochemie; CB - Analytická chemie, separace (UIACH-O)
Obor OECD: Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis); Analytical chemistry (UIACH-O)
Impakt faktor: 5.116, rok: 2017
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.
Trvalý link: http://hdl.handle.net/11104/0272786