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

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F68081707%3A_____%2F17%3A00476223" target="_blank" >RIV/68081707:_____/17:00476223 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/68081715:_____/17:00476223

Výsledek na webu

<a href="http://dx.doi.org/10.1016/j.electacta.2017.04.045" target="_blank" >http://dx.doi.org/10.1016/j.electacta.2017.04.045</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1016/j.electacta.2017.04.045" target="_blank" >10.1016/j.electacta.2017.04.045</a>

Jazyk výsledku

angličtina

Název v původním jazyce

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

Popis výsledku v původním jazyce

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.

Název v anglickém jazyce

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

Popis výsledku anglicky

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.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

—

OECD FORD obor

10405 - Electrochemistry (dry cells, batteries, fuel cells, corrosion metals, electrolysis)

Projekt

<a href="/cs/project/GA15-15479S" target="_blank" >GA15-15479S: Nové nástroje pro výzkum a diagnostiku nemocí. Mikrofluidické reaktory a elektrochemie pro analýzu proteinů a jejich glykosylaci.</a><br>

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2017

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Electrochimica acta

ISSN

0013-4686

e-ISSN

—

Svazek periodika

239

Číslo periodika v rámci svazku

JUN 2017

Stát vydavatele periodika

GB - Spojené království Velké Británie a Severního Irska

Počet stran výsledku

6

Strana od-do

10-15

Kód UT WoS článku

000401114000002

EID výsledku v databázi Scopus

—