Functional delivery of a membrane protein into oocyte membranes using bicelles.
Voltage-gated potassium channel modulatory membrane protein KCNE3 was overexpressed and purified into both micelles and bicelles. Remarkably, microinjection of KCNE3 in bicelles into Xenopus oocytes resulted in functional co-assembly with the human KCNQ1 channel expressed therein. Microinjection of LMPC micelles containing KCNE3 did not result in channel modulation, indicating that bicelles sometimes succeed at delivering a membrane protein into a cellular membrane when classical micelles fail. Backbone NMR resonance assignments were completed for KCNE3 in both bicelles and LMPC, indicating that the secondary structure distribution in KCNE3's N-terminus and transmembrane domains exhibits only modest differences from that of KCNE1, even though these KCNE family members have very different effects on KCNQ1 channel function.
Kang
Congbao
C
Department of Biochemistry, Center for Structural Biology, Vanderbilt University School of Medicine, Nashville, Tennessee 37232-8725, USA.
Vanoye
Carlos G
CG
Welch
Richard C
RC
Van Horn
Wade D
WD
Sanders
Charles R
CR
D000595:Amino Acid Sequence; D000818:Animals; D006801:Humans; D051657:KCNQ1 Potassium Channel / Q000737:chemistry / Q000378:metabolism; D008564:Membrane Potentials; D008565:Membrane Proteins / Q000737:chemistry / Q000378:metabolism*; D008823:Micelles; D008845:Microinjections; D008954:Models, Biological; D008969:Molecular Sequence Data; D009865:Oocytes / Q000378:metabolism*; D024642:Potassium Channels, Voltage-Gated / Q000737:chemistry / Q000378:metabolism*; D014981:Xenopus / Q000378:metabolism
C418492:KCNE3 protein, human; D051657:KCNQ1 Potassium Channel; C496909:KCNQ1 protein, human; D008565:Membrane Proteins; D008823:Micelles; D024642:Potassium Channels, Voltage-Gated