Carnarius, Christian

Connexin 43 (Cx43) is the most prominent
member of the connexin family in mammalian tissues. In biological
lipid membranes, six connexin subunits assemble into a hemichannel
(connexon). Direct communication of apposing cells is realized by
two adjacent hemichannels, forming gap junction channels. Membrane
channel proteins are pivotal for communication between cells.
Dysfunctional ion channels are associated to diseases like
osteoporosis, epilepsy, ataxia, myotonia and cardiac arrhythmia.
Therefore, there is an interest in studying these channel proteins
to elucidate their functional properties and determine how to
manipulate their functionality. This is commonly carried out by ion
channel recordings, which require invasive contacting by voltage
clamp methods in cells. The alternative approach taken here was to
utilize connexon-doped artificial membranes for electrical coupling
of connexon-expressing cells via gap junction channels. In this
work, an expression system in Pichia pastoris to recombinantly
produce and purify Cx43 as well as Cx43 fused to green fluorescent
protein (Cx4

Functional reconstitution of connexons in artificial membranes: expression, purification and characterization of connexin 43

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Georg-August-University Göttingen