Native Planar Asymmetric Suspended Membrane for Single‐Molecule Investigations: Plasma Membrane on a Chip. Issue 51 (3rd November 2022)

• Record Type: Journal Article
• Authors: Kalyana Sundaram, Ramalingam Venkat; Bera, Manindra; Coleman, Jeff; Weerakkody, Jonathan S.; Krishnakumar, Shyam S.; Ramakrishnan, Sathish
• Is Part Of: Small. Volume 18:Issue 51(2022)
• Page Start: n/a
• Page End: n/a
• Publication Date: 2022-11-03
• Subjects: asymmetric lipid membranes -- native suspended plasma membranes -- protein diffusion -- protein fluorescent resonant energy transfer (FRET) -- silicon microarrays -- single molecule counting -- total internal reflection fluorescence penetration depth; Nanotechnology -- Periodicals; Nanoparticles -- Periodicals; Microtechnology -- Periodicals; 620.5
• DOI: 10.1002/smll.202205567
• Languages: English
• ISSNs: 1613-6810

Abstract

Abstract: Cellular plasma membranes, in their role as gatekeepers to the external environment, host numerous protein assemblies and lipid domains that manage the movement of molecules into and out of cells, regulate electric potential, and direct cell signaling. The ability to investigate these roles on the bilayer at a single‐molecule level in a controlled, in vitro environment while preserving lipid and protein architectures will provide deeper insights into how the plasma membrane works. A tunable silicon microarray platform that supports stable, planar, and asymmetric suspended lipid membranes (SLIM) using synthetic and native plasma membrane vesicles for single‐molecule fluorescence investigations is developed. Essentially, a "plasma membrane‐on‐a‐chip" system that preserves lipid asymmetry and protein orientation is created. By harnessing the combined potential of this platform with total internal reflection fluorescence (TIRF) microscopy, the authors are able to visualize protein complexes with single‐molecule precision. This technology has widespread applications in biological processes that happen at the cellular membranes and will further the knowledge of lipid and protein assemblies. Abstract : A tunable silicon‐based platform is presented that supports suspended lipid membranes (SLIM) to interrogate membrane protein assembly and dynamics using total internal reflection fluorescence (TIRF) microscopy. By generating bilayers from giant plasma membrane vesicles (GPMVs) derived from different cell types, fast dynamics of protein function on the inner leaflet of cells are captured while retaining native lipid architecture and asymmetry.