Electrochemical Platform for the Detection of Transmembrane Proteins Reconstituted into Liposomes

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F16%3AN0000136" target="_blank" >RIV/00027162:_____/16:N0000136 - isvavai.cz</a>

Alternative codes found

RIV/61989592:15310/16:33159000 RIV/61989592:15110/16:33159000 RIV/68378271:_____/16:00478799

Result on the web

<a href="http://pubs.acs.org/doi/abs/10.1021/acs.analchem.6b00618" target="_blank" >http://pubs.acs.org/doi/abs/10.1021/acs.analchem.6b00618</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acs.analchem.6b00618" target="_blank" >10.1021/acs.analchem.6b00618</a>

Result language

angličtina

Original language name

Electrochemical Platform for the Detection of Transmembrane Proteins Reconstituted into Liposomes

Original language description

The development of new methods and strategies for the investigation of membrane proteins is limited by poor solubility of these proteins in an aqueous environment and hindered by a number of other problems linked to the instability of the proteins outside lipid bilayers. Therefore, current research focuses on an analysis of membrane proteins incorporated into model lipid membrane, most frequently liposomes. In this work, we introduce a new electrochemical methodology for the analysis of transmembrane proteins reconstituted into a liposomal system. The proposed analytical approach is based on proteoliposomal sample adsorption on the surface of working electrodes followed by analysis of the anodic and cathodic signals of the reconstituted proteins. It works based on the fact that proteins are electroactive species, in contrast to the lipid components of the membranes under the given experimental conditions. Electroanalytical experiments were performed with two transmembrane proteins; the Na+/K+ATPase that contains transmembrane as well as large extramembraneous segments and the mitochondrial uncoupling protein 1, which is a transmembrane protein essentially lacking extramembraneous segments. Electrochemical analyses of proteoliposomes were compared with analyses of both proteins solubilized with detergents (C12E8 and octyl-PoE) and supported by the following complementary methods: microscopy techniques, protein activity testing, molecular model visualizations, and immunochemical identification of both proteins. The label-free electrochemical platform presented here enables studies of reconstituted transmembrane proteins at the nanomolar level. Our results may contribute to the development of new electrochemical sensors and microarray systems applicable within the field of poorly water-soluble proteins.

Czech name

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Czech description

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Type

J_x - Unclassified - Peer-reviewed scientific article (Jimp, Jsc and Jost)

CEP classification

CB - Analytical chemistry, separation

OECD FORD branch

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Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)<br>I - Institucionalni podpora na dlouhodoby koncepcni rozvoj vyzkumne organizace

Publication year

2016

Confidentiality

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

Name of the periodical

Analytical Chemistry

ISSN

0003-2700

e-ISSN

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Volume of the periodical

88

Issue of the periodical within the volume

8

Country of publishing house

US - UNITED STATES

Number of pages

9

Pages from-to

4548-4556

UT code for WoS article

000374706000052

EID of the result in the Scopus database

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