Analysis of membrane systems using lipoprotein nanodiscs

Membrane systems play a critical role in biochemistry by regulating the chemical, energy, and information flow into the cell and its various compartments. However, the amphipathic nature of membrane proteins and lipids makes their analysis challenging. Engineered nanoscale lipoprotein Nanodiscs provide a well-defined and native-like lipid bilayer to solubilize membrane proteins and lipids. This dissertation details the analysis of Nanodiscs containing various membrane proteins and lipids with bioanalytical methods, including mass spectrometry and photonic biosensors. Mass spectrometry offers several powerful techniques for fundamental characterization of Nanodiscs and Nanodisc-solubilized membrane proteins. Native mass spectrometry delivers single-lipid resolution of intact Nanodisc complexes to determine their mass distributions. Mass spectrometry-based proteomics demonstrates that heterogeneous Nanodisc-solubilized membrane protein libraries model the membrane proteome. Matrix-associated laser desorption ionization methods facilitate characterization of membrane proteins embedded in Nanodiscs. To study biomolecular interactions involving membrane systems, silicon photonic microring resonator biosensors provide a multiplexed platform for detecting interactions between soluble proteins and Nanodiscs. A novel direct physisorption strategy was developed to interface Nanodiscs with the silica surface. Implementation of a nonlinear analyte gradient approach extends the capabilities of the microring resonator technology to single run kinetic experiments. Mass spectrometry and silicon photonic biosensor techniques developed in this thesis extend the capabilities of the Nanodisc platform for the analysis of membrane systems and lay the foundation for future applications of Nanodiscs in biochemistry, target identification, and drug discovery.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Chemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2014

Author dc:creator

• Marty, Michael

Contributors dc:contributor

• Sligar, Stephen G.
• Sweedler, Jonathan V.
• Bailey, Ryan C.
• Kraft, Mary L.

• Nanodiscs
• Lipoprotein Particles
• Membrane Proteins
• Lipids
• Mass Spectrometry
• Photonic Biosensors
• Proteomics

Statement dc:rights

• Copyright 2013 Michael T. Marty

Language dc:language

en

Handle dc:identifier

http://hdl.handle.net/2142/46869

OAI identifier oai:identifier

oai:www.ideals.illinois.edu:2142/46869