Soloviev, Z;
(2017)
Structural mass spectrometry studies of proteins and protein complexes involved in the regulation of gene expression.
Doctoral thesis , UCL (University College London).
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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.
| Type: | Thesis (Doctoral) |
|---|---|
| Title: | Structural mass spectrometry studies of proteins and protein complexes involved in the regulation of gene expression |
| Event: | UCL (University College London) |
| Open access status: | An open access version is available from UCL Discovery |
| Language: | English |
| UCL classification: | UCL UCL > Provost and Vice Provost Offices UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Div of Biosciences > Structural and Molecular Biology |
| URI: | https://discovery.ucl.ac.uk/id/eprint/1541272 |
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