title: In vitro reconstitution of septin assemblies on supported lipid bilayers.
abstract: Septins are polymerizing eukaryotic proteins that play conserved roles in cell cortex organization and are essential in many cell types. How septin dynamics and protein-protein interactions determine their function at the plasma membrane remains a mystery. Here, we present a method for recapitulating septin polymerization and lipid interaction utilizing supported lipid bilayers to mimic the eukaryotic plasma membrane. Septins on supported lipid bilayers can be visualized with single-molecule sensitivity using total internal reflective fluorescence microscopy. Microscopy-based in vitro assays have revolutionized our understanding of actin, microtubules, and bacterial cytoskeletal systems, and will likely immediately advance our understanding of the septin proteins. As such, we hope that this technique will be adopted and widely utilized by those interested in uncovering septin properties and functions of septin interacting proteins.
authors.affiliation: Dartmouth College, Hanover, NH, United States; Marine Biological Laboratory, Woods Hole, MA, United States.
authors.forename: A A
authors.initials: AA
authors.lastname: Bridges
authors.affiliation: Dartmouth College, Hanover, NH, United States; Marine Biological Laboratory, Woods Hole, MA, United States.
authors.forename: A S
authors.initials: AS
authors.lastname: Gladfelter
mesh_terms: D000199:Actins / Q000737:chemistry; D001419:Bacteria / Q000737:chemistry / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry / Q000648:ultrastructure; D003593:Cytoplasm / Q000737:chemistry / Q000648:ultrastructure; D003599:Cytoskeleton / Q000648:ultrastructure; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D008853:Microscopy / Q000379:methods*; D008870:Microtubules / Q000737:chemistry / Q000648:ultrastructure*; D060066:Protein Interaction Maps; D055503:Protein Multimerization; D058112:Septins / Q000737:chemistry* / Q000648:ultrastructure
chemical_list: D000199:Actins; D008051:Lipid Bilayers; D058112:Septins
keywords: Cytoskeleton; In vitro; Reconstitution; Septin; Supported lipid bilayer