PubMed 7694656

Referenced in Channelpedia wiki pages of: none

Automatically associated channels: Kv1.5 , Slo1

Title: Functional and biochemical characterization of the human potassium channel Kv1.5 with a transplanted carboxyl-terminal epitope in stable mammalian cell lines.

Authors: L H Philipson, A Malayev, A Kuznetsov, C Chang, D J Nelson

Journal, date & volume: Biochim. Biophys. Acta, 1993 Nov 21 , 1153, 111-21

PubMed link:

The role of the C-terminal domain of the hPCN1/Kv1.5 delayed rectifier K+ channel was investigated in transfected stable cell lines employing antipeptide and anti-epitope antibodies against hPCN1-cp, an epitope-fusion gene carrying additional sequences encoding a 32 amino acid C-terminal extension. Both wild-type and chimeric genes showed high levels of K+ channel expression. Detailed electrophysiologic characterization showed there to be no significant effect of the C-terminal extension on channel activity. Immunoblots of whole-cell and membrane preparations demonstrated primarily intact protein in which the C-terminal extension was not cleaved from the peptide chain. Two bands were visualized from cells transfected with either the wild-type or chimeric channels; the slower migrating band was a non-N-glycosylated form. The epitope-fusion method will be a useful adjunct to studying the role of functional domains in ion channels, and may provide a means for rapid affinity purification of channel protein.