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Title: Studying protein-ligand complexes in the gas-phase using ion mobility-mass spectrometry
Author: Hopper, Jonathan T. S.
ISNI:       0000 0004 2742 5885
Awarding Body: University of Nottingham
Current Institution: University of Nottingham
Date of Award: 2012
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This thesis presents studies which investigate the application of electrospray ionisation-mass spectrometry (ESI-MS) to non-covalent protein-ligand complexes. The structural effect of desolvation on protein-ligand (P-L) complexes has been a hotly debated issue in this field and has been examined in this work. Single point mutagenesis has allowed specific non-covalent interactions to be probed as well as their contribution to gas-phase protein- ligand stability. Results suggest that these specific interactions are preserved in the gas-phase using ESI. Other solution based effects that result from ligand binding, such as increased protein structural stability, was also confirmed in the gas-phase. A combination of collisional activation and ion mobility spectrometry is presented as an approach capable of probing such subtle stability differences. Some discrepancies between the behaviour of protein- ligand complexes in solution and the gas-phase are also presented and highlight potential areas of caution in certain biological systems. Common alkali metal adducts have been shown to severely decrease the stability of protein-ligand complexes in the gas-phase, possibly by a Coulomb assisted dissociation mechanism. Novel approaches to allow greater control of charge state distributions, without the requirement of instrumental modifications, are also presented. Reducing the charge state of protein complexes in the gas-phase allows weak interactions to be more readily preserved and more accurate affinity measurements to be made. The approach is also confirmed to reduce the amount of alkali metal adduction observed in protein ions generated by ESI.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available