Use this URL to cite or link to this record in EThOS: https://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.790690
Title: Structural mass spectrometry studies of proteins and protein complexes involved in the regulation of gene expression
Author: Soloviev, Z.
ISNI:       0000 0004 8498 8668
Awarding Body: UCL (University College London)
Current Institution: University College London (University of London)
Date of Award: 2017
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Abstract:
Mass spectrometry is becoming an ever more popular technique of choice for studying protein structure and dynamics. This thesis describes multiple uses of mass spectrometry and its applications in the field of structural biology. The domain interaction and function of the Human Histone Deacetylase 2 (HDAC2) protein is covered in Chapters 2 and 3, where work performed ranged from protein expression and purification to mass spectrometry, activity analysis and molecular modelling. Activity assays together with intact mass analysis and cross-linking mass spectrometry allowed to dissect the interaction between the intrinsically-disorderend domain and the active site of the protein, where the intrinsically-disordered domain's flexibility allows to inhibit protein's function. Charge manipulation experiments and the importance of careful methods development is described in Chapter 4, where two proteins of different physical and chemical properties were subjected to charge altering buffer conditions and analysed using native mass spectrometry, collision-induced unfolding, and top-down fragmentation, which showed that flexible and globular proteins react differently to charge altering conditions Finally, Chapter 5 covers the study of two different metal-binding proteins and the structural re-arrangements induced upon metal binding. Native mass spectrometry was used alongside ion mobility mass spectrometry and collision-induced unfolding to observe changes in protein stability upon metal ion binding. In both cases, metal ions showed a stabilising effect on the protein structure. Specificity of metal binding was observed by investigating mutant proteins, which failed to bind metal ions with the same affinity as the wilt-type protein.
Supervisor: Thalassinos, K. ; Hansen, D. Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID: uk.bl.ethos.790690  DOI: Not available
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