Fractional polymerization of a suspended planar bilayer creates a fluid, highly stable membrane for ion channel recordings.
Suspended planar lipid membranes (or black lipid membranes (BLMs)) are widely used for studying reconstituted ion channels, although they lack the chemical and mechanical stability needed for incorporation into high-throughput biosensors and biochips. Lipid polymerization enhances BLM stability but is incompatible with ion channel function when membrane fluidity is required. Here, we demonstrate the preparation of a highly stable BLM that retains significant fluidity by using a mixture of polymerizable and nonpolymerizable phospholipids. Alamethicin, a voltage-gated peptide channel for which membrane fluidity is required for activity, was reconstituted into mixed BLMs prepared using bis-dienoyl phosphatidylcholine (bis-DenPC) and diphytanoyl phosphatidylcholine (DPhPC). Polymerization yielded BLMs that retain the fluidity required for alamethicin activity yet are stable for several days as compared to a few hours prior to polymerization. Thus, these polymerized, binary composition BLMs feature both fluidity and long-term stability.
Heitz
Benjamin A
BA
Department of Chemistry and Biochemistry and BIO5 Institute, University of Arizona, Tucson, Arizona 85721, USA.
Jones
Ian W
IW
Hall
Henry K
HK
Aspinwall
Craig A
CA
Saavedra
S Scott
SS
D000408:Alamethicin / Q000378:metabolism; D007473:Ion Channels / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008560:Membrane Fluidity; D008567:Membranes, Artificial; D010713:Phosphatidylcholines / Q000737:chemistry / Q000378:metabolism; D011108:Polymers / Q000737:chemistry* / Q000378:metabolism*; D013535:Suspensions; D013997:Time Factors
D007473:Ion Channels; D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010713:Phosphatidylcholines; D011108:Polymers; D013535:Suspensions; D000408:Alamethicin