Rapid microfluidic perfusion enabling kinetic studies of lipid ion channels in a bilayer lipid membrane chip.

ABSTRACT: There is growing recognition that lipids play key roles in ion channel physiology, both through the dynamic formation and dissolution of lipid ion channels and by indirect regulation of protein ion channels. Because existing technologies cannot rapidly modulate the local (bio)chemical conditions at artificial bilayer lipid membranes used in ion channel studies, the ability to elucidate the dynamics of these lipid-lipid and lipid-protein interactions has been limited. Here we demonstrate a microfluidic system supporting exceptionally rapid perfusion of reagents to an on-chip bilayer lipid membrane, enabling the responses of lipid ion channels to dynamic changes in membrane boundary conditions to be probed. The thermoplastic microfluidic system allows initial perfusion of reagents to the membrane in less than 1 s, and enables kinetic behaviors with time constants below 10 s to be directly measured. Application of the platform is demonstrated toward kinetic studies of ceramide, a biologically important lipid known to self-assemble into transmembrane ion channels, in response to dynamic treatments of small ions (La(3+)) and proteins (Bcl-x(L) mutant). The results reveal the broader potential of the technology for studies of membrane biophysics, including lipid ion channel dynamics, lipid-protein interactions, and the regulation of protein ion channels by lipid micro domains.

SUBMITTER: Shao C 

PROVIDER: S-EPMC3343723 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature