Open AccessSupported Lipid Bilayer Technology for the Study of Cellular Interfaces
Author(s) -
Crites Travis J.,
Maddox Michael,
Padhan Kartika,
Muller James,
Eigsti Calvin,
Varma Rajat
Publication year - 2015
Publication title -
current protocols in cell biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.149
H-Index - 38
eISSN - 1934-2616
pISSN - 1934-2500
DOI - 10.1002/0471143030.cb2405s68
Subject(s) - jurkat cells , microbiology and biotechnology , liposome , lipid bilayer , flow cytometry , immunological synapse , cell , transmembrane protein , population , chemistry , biology , immune system , t cell , membrane , biochemistry , immunology , t cell receptor , receptor , demography , sociology
Glass‐supported lipid bilayers presenting freely diffusing proteins have served as a powerful tool for studying cell‐cell interfaces, in particular, T cell–antigen presenting cell (APC) interactions, using optical microscopy. Here we expand upon existing protocols and describe the preparation of liposomes by an extrusion method, and describe how this system can be used to study immune synapse formation by Jurkat cells. We also present a method for forming such lipid bilayers on silica beads for the study of signaling responses by population methods, such as western blotting, flow cytometry, and gene‐expression analysis. Finally, we describe how to design and prepare transmembrane‐anchored protein‐laden liposomes, following expression in suspension CHO (CHOs) cells, a mammalian expression system alternative to insect and bacterial cell lines, which do not produce mammalian glycosylation patterns. Such transmembrane‐anchored proteins may have many novel applications in cell biology and immunology. © 2015 by John Wiley & Sons, Inc.