Synthetic Ion Channels: From Pores to Biological Applications

In this Account, we describe the development of several diverse families I of synthetic, membrane-active amphiphiles that form pores and facilitate transport within membrane bilayers. For the most part, the compounds are amphiphiles that insert into the bilayer and form pores either on their own or by self-assembly. The first family of synthetic ion channels prepared in our lab, the hydraphiles, used crown ethers as head groups and as a polar central element. In a range of biophysical studies, we showed that the hydraphiles formed unimolecular pores that spanned the bilayer. They mediated the transport of Na+ and K+ but were blocked by Ag+. The hydraphiles are nonrectifying and disrupt ion homeostasis. As a result, these synthetic ion channels are toxic to various bacteria and yeast, a feature that has been used therapeutically in direct injection chemotherapy. We also developed a family of amphiphilic heptapeptide ion transporters that selected Cl- >10-fold over K+ and showed voltage dependent gating. The formed pores were approximately dimeric, and variations in the N- and C-terminal anchor chains and the adds affected transport rates. Surprisingly, the longer N-terminal anchor chains led to less transport but greater Cl- selectivity. A proline residue, which is present in the CIC protein channel's conductance pore, proved to be critical for Cl- transport selectivity. Pyrogallol[4]arenes are macrocydes formed by acid-catalyzed condensation of four 1,2,3- trihydroxybenzenes with four aldehydes. The combination of 12 hydroxyl groups on one face of the macrocycle and four pendant alkyl chains conferred considerable amphiphilicity to these compounds. The pyrogallol[4]arenes inserted into bilayer membranes and conducted ions. Based on our experimental evidence, the ions passed through a self-assembled pore comprising four or five amphiphiles rather than passing through the central opening of a single macrocycle. Pyrogallol[4]arenes constructed with branched chains are also amphiphilic and active in membranes. The pyrogallol[4]arene with 3-pentyl sidechains formed a unique nanotube assembly and functioned as an ion channel in bilayer membranes. Finally, we showed that dianilides of either isophthalic or dipicolinic acids, compounds which have been extensively studied as anion binders, can self-assemble to form pores within bilayers. We called these dianilides tris-arenes and have shown that they readily bind to phosphate anions. These structures also mediated the transport of DNA plasmids through vital bilayer membranes in the bacterium Escherichia coli and in the yeast Saccharomyces cerevisiae. This transformation or transfection process occurred readily and without any apparent toxicity or mutagenicity.