Open AccessApplications of Lipid Nanodiscs for the Study of Membrane Proteins by Surface Plasmon Resonance
Author(s) -
Trahey Meg,
Li Mavis Jiarong,
Kwon Hyewon,
Woodahl Erica L.,
McClary Wynton D.,
Atkins William M.
Publication year - 2015
Publication title -
current protocols in protein science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.409
H-Index - 32
eISSN - 1934-3663
pISSN - 1934-3655
DOI - 10.1002/0471140864.ps2913s81
Subject(s) - surface plasmon resonance , polyclonal antibodies , nanodisc , chemistry , lipid bilayer , membrane , nanotechnology , membrane protein , biophysics , materials science , antibody , biochemistry , biology , nanoparticle , immunology
Methods for the initial steps of surface plasmon resonance analysis of membrane proteins incorporated in lipid nanodiscs are described. Several types of Biacore sensor chips are available and require distinct strategies to immobilize proteonanodiscs on the chip surface. The procedures for immobilization on three of these chips (NTA, antibody coupled CM5, and L1) are described in this unit and results are demonstrated for a model system with cytochrome P4503A4 (CYP3A4) in nanodiscs binding to a polyclonal anti‐CYP3A4 antibody. Advantages and disadvantages of each chip type are considered. © 2015 by John Wiley & Sons, Inc.