Functional hemichannels formed by human connexin 26 expressed in bacteria
Journal
Bioscience Reports
ISSN
0144-8463
1573-4935
Date Issued
2015
Author(s)
Mariana C. Fiori
Srinivasan Krishnan
D. Marien Cortes
Luis Reuss
Guillermo A. Altenberg
Luis G. Cuello
Type
Resource Types::text::journal::journal article
URL Institutional Repository
Abstract
<jats:p>Gap-junction channels (GJCs) communicate the cytoplasm of adjacent cells and are formed by head-to-head association of two hemichannels (HCs), one from each of the neighbouring cells. GJCs mediate electrical and chemical communication between cells, whereas undocked HCs participate in paracrine signalling because of their permeability to molecules such as ATP. Sustained opening of HCs under pathological conditions results in water and solute fluxes that cannot be compensated by membrane transport and therefore lead to cell damage. Mutations of Cx26 (connexin 26) are the most frequent cause of genetic deafness and it is therefore important to understand the structure–function relationship of wild-type and deafness-associated mutants. Currently available connexin HC expression systems severely limit the pace of structural studies and there is no simple high-throughput HC functional assay. The Escherichia coli-based expression system presented in the present study yields milligram amounts of purified Cx26 HCs suitable for functional and structural studies. We also show evidence of functional activity of recombinant Cx26 HCs in intact bacteria using a new growth complementation assay. The E. coli-based expression system has high potential for structural studies and high-throughput functional screening of HCs.</jats:p>
Subjects
adenosine 5 '-triphosphate (atp)
;
calcium
;
connexon
;
deafness
;
gap-junction channel
;
method