Universität zu Köln

Perniss, Alexander, Wolf, Annemarie, Wichmann, Lukas, Schoenberger, Matthias and Althaus, Mike ORCID: 0000-0002-3554-7538 (2015). Evans Blue is not a suitable inhibitor of the epithelial sodium channel delta-subunit. Biochem. Biophys. Res. Commun., 466 (3). S. 468 - 475. SAN DIEGO: ACADEMIC PRESS INC ELSEVIER SCIENCE. ISSN 1090-2104

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Abstract

The Epithelial Sodium Channel (ENaC) is a heterotrimeric ion channel which can be either formed by assembly of its alpha-, beta- and gamma-subunits or, alternatively, its delta-, beta- and gamma-subunits. The physiological function of a alpha beta gamma-ENaC is well established, but the function of delta beta gamma-ENaC remains elusive. The azo-dye Evans Blue (EvB) has been routinely used to discriminate between the two channel isoforms by decreasing transmembrane currents and amiloride-sensitive current fractions of delta beta gamma-ENaC expressing Xenopus oocytes. Even though these results could be reproduced, it was found by precipitation experiments and spectroscopic methods that the cationic amiloride and the anionic EvB directly interact in solution, forming a strong complex. Thereby a large amount of pharmacologically available amiloride is removed from physiological buffer solutions and the effective amiloride concentration is reduced. This interaction did not occur in the presence of albumin. In microelectrode recordings, EvB was able to abrogate the block of delta beta gamma-ENaC by amiloride or its derivative benzamil. In sum, EvB reduces amiloride-sensitive ion current fractions in electrophysiological experiments. This is not a result of a specific inhibition of delta beta gamma-ENaC but rather represents a pharmacological artefact. EvB should therefore not be used as an inhibitor of delta-ENaC. (C) 2015 Elsevier Inc. All rights reserved.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Perniss, Alexander UNSPECIFIED UNSPECIFIED UNSPECIFIED Wolf, Annemarie UNSPECIFIED UNSPECIFIED UNSPECIFIED Wichmann, Lukas UNSPECIFIED UNSPECIFIED UNSPECIFIED Schoenberger, Matthias UNSPECIFIED UNSPECIFIED UNSPECIFIED Althaus, Mike UNSPECIFIED orcid.org/0000-0002-3554-7538 UNSPECIFIED
URN:	urn:nbn:de:hbz:38-389424
DOI:	10.1016/j.bbrc.2015.09.052
Journal or Publication Title:	Biochem. Biophys. Res. Commun.
Volume:	466
Number:	3
Page Range:	S. 468 - 475
Date:	2015
Publisher:	ACADEMIC PRESS INC ELSEVIER SCIENCE
Place of Publication:	SAN DIEGO
ISSN:	1090-2104
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language NA+ CHANNEL; AMILORIDE; ENAC Multiple languages Biochemistry & Molecular Biology; Biophysics Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/38942