title: Use of the supported membrane tube assay system for real-time analysis of membrane fission reactions.
abstract: The process of membrane fission is fundamental to diverse cellular processes such as nutrient uptake, synaptic transmission and organelle biogenesis, and it involves the localized application of curvature stress to a tubular membrane intermediate, forcing it to undergo scission. Alternative techniques for creating such substrates necessitate the use of micromanipulators or sophisticated optical traps and require a high level of technical expertise. We present a facile method to generate an array of membrane tubes supported on a passivated glass coverslip, which we refer to as supported membrane tubes (SMrTs). SMrT templates are formed upon hydration of a dry lipid mix in physiological buffer and subsequent flow-induced extrusion of the lipid reservoir into long membrane tubes with variable dimensions. Following surface passivation of coverslips, these templates can be formed from a variety of lipids, with as little as 1-2 nmol of lipid in a matter of 2 h, and can be used in membrane-curvature-sensitive fission assays.
authors.affiliation: Indian Institute of Science Education and Research, Pune, Maharashtra, India.
authors.forename: Srishti
authors.initials: S
authors.lastname: Dar
authors.affiliation: Indian Institute of Science Education and Research, Pune, Maharashtra, India.
authors.forename: Sukrut C
authors.initials: SC
authors.lastname: Kamerkar
authors.affiliation: Indian Institute of Science Education and Research, Pune, Maharashtra, India.
authors.forename: Thomas J
authors.initials: TJ
authors.lastname: Pucadyil
mesh_terms: D002021:Buffers; D002462:Cell Membrane / Q000378:metabolism*; D005898:Glass / Q000737:chemistry; D050356:Lipid Metabolism; D008567:Membranes, Artificial; D013997:Time Factors
chemical_list: D002021:Buffers; D008567:Membranes, Artificial