Incorporation and analysis of bilayer constituents in nanodiscs

Nanodiscs, engineered lipoparticles composed of a lipid bilayer and two surrounding amphipathic proteins, have been utilized to house and solubilize integral membrane proteins and other molecules with large hydrophobic groups which limit their aqueous solubility. In the experiments described herein, lipids from the Nanodiscs’ bilayer, gadolinium chelates and integral membrane proteins were analyzed for their incorporation and stability within the Nanodisc structure. Four techniques are described for quantification of the exchange of lipids between Nanodiscs over time, including: scintillation counting, imaging proximity assay, NMR and TLC MALDI-MS. Additionally, the extent and longevity of inclusion of the hydrophobic gadolinium-chelate “C3lipo” doped into the Nanodisc bilayer was analyzed and preliminary cell uptake studies were performed. Finally, the incorporation of proteins from rat liver and fetal bovine brain membranes into Nanodiscs to create soluble membrane protein libraries is discussed.

Name thesis:degree_name

M.S.

Level thesis:degree_level

Thesis

Discipline thesis:degree_discipline

Chemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Baker, Catherine

Contributors dc:contributor

• Sligar, Stephen G.

• Nanodiscs
• Lipid exchange
• Soluble membrane protein library (SMPL)
• Lipoprotein
• Membrane scaffold protein
• Gadolinium

Statement dc:rights

• Copyright 2013 Catherine Baker

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/46921

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/46921