Analysis of membrane systems using lipoprotein nanodiscs

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

Marty, Michael

关键词: Nanodiscs;    Lipoprotein Particles;    Membrane Proteins;    Lipids;    Mass Spectrometry;    Photonic Biosensors;    Proteomics;   

Others  :  https://www.ideals.illinois.edu/bitstream/handle/2142/46869/Michael_Marty.pdf?sequence=1&isAllowed=y

美国|英语

来源: The Illinois Digital Environment for Access to Learning and Scholarship

【 摘 要 】

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.