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Complementary Mass Spectrometry Methods For Characterizing Protein Folding, Structure, And Dynamics, Siavash Vahidi
Complementary Mass Spectrometry Methods For Characterizing Protein Folding, Structure, And Dynamics, Siavash Vahidi
Electronic Thesis and Dissertation Repository
Proteins are involved in virtually every biochemical process. A comprehensive characterization of factors that govern protein function is essential for understanding the biomedical aspects of human health. This dissertation aims to develop complementary mass spectrometry-based methods and apply them to solve problems pertaining to the area of protein structure, folding and dynamics.
Chapter 1 uses fast photochemical oxidation of proteins (FPOP) to characterize partially disordered conformers populated under semi-denaturing conditions. In FPOP, ·OH generated by laser photolysis of H2O2 introduces oxidative modifications at solvent accessible side chains. By contrast, buried sites are protected from radical attack. Using …
Structure And Dynamics Of The Membrane Protein Bacteriorhodopsin Studied By Mass Spectrometry, Yan Pan
Structure And Dynamics Of The Membrane Protein Bacteriorhodopsin Studied By Mass Spectrometry, Yan Pan
Electronic Thesis and Dissertation Repository
Membrane proteins continue to represent a major challenge for most analytical techniques. Using bacteriorhodopsin (BR) as model system, this work aims to develop mass spectrometry (MS)-based approaches for exploring the structure, dynamics and folding of membrane proteins.
As the first step, BR in its native lipid environment was exposed to hydroxyl radicals, which were produced by laser photolysis of hydrogen peroxide. It was found that the resulting methionine (Met) labeling pattern was consistent with the known BR structure. This finding demonstrates that laser-induced oxidative Met labeling can provide structural information on membrane proteins. In subsequent experiments, the effects of different …