Abstract
KCNE1, also known as MinK, is a membrane protein that associates with the KCNQ1 channel protein to form a voltage-gated potassium channel. This ion channel is essential to the cardiac action potential that mediates heartbeat and is also critical for potassium ion homeostasis in the inner ear. Dominant mutations in KCNE1 lead to congenital long-QT syndrome and congenital deafness. KCNE1 has been over expressed in E. coli, purified into micelles using his-tag affinity chromatography, and reconstituted into POPC/POPG vesicles. 31P NMR powder spectra results confirm vesicle formation. Different KCNE1 mutants have been labeled using MTSL, one mutant outside the membrane and the other inside the membrane. By measuring 31P relaxation times of the lipids, we can determine the depth that at which KCNE1 is buried inside the vesicles. We also introduced a bicelle system to study the topology of uniform 15N labeled KCNE1 with respect to the lipid bilayer. By measuring the 15N NMR signal, we are able to figure out the structural topology of KCNE1 within the lipid bilayer.
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