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dc.contributor.advisorBarran, Perdita
dc.contributor.advisorBarlow, Paul
dc.contributor.authorJurneczko, Ewa
dc.date.accessioned2014-05-23T13:38:14Z
dc.date.available2014-05-23T13:38:14Z
dc.date.issued2014-06-28
dc.identifier.urihttp://hdl.handle.net/1842/8844
dc.description.abstractFor proteins the link between their structure and their function is a central tenet of biology. A common approach to understanding protein function is to ‘solve’ its structure and subsequently probe interactions between the protein and its binding partners. The first part of this approach is non-trivial for proteins where localised regions or even their entire structure fail to fold into a three-dimensional structure and yet they possess function. These so called intrinsically or inherently disordered proteins (IDP’s) or intrinsically disordered regions (IDR’s) constitute up to 40% of all expressed proteins. IDPs which have crucial roles in molecular recognition, assembly, protein modification and entropic chain activities, are often dynamic with respect to both conformation and interaction, so in the course of a protein’s ‘lifespan’ it will sample many configurations and bind to several targets. For these proteins, there is a need to develop new methods for structure characterization which exploit their biophysical properties. The solvent free environment of a mass spectrometer is ideally suited to the study of intrinsic interactions and how they contribute to structure. Ion mobility mass spectrometry is uniquely able to observe the range of structures an IDP can occupy, and also the effect of selected binding partners on altering this conformational space. This thesis details the technique of ion mobility mass spectrometry and illustrates its use in assessing the relative disorder of p53 protein. The tumour suppressor p53 is at the hub of a plethora of signalling pathways that maintain the integrity of the human genome and regulate the cell cycle. Deregulation of this protein has a great effect on carcinogenesis as mutated p53 can induce an amplified epigenetic instability of tumour cells, facilitating and accelerating the evolution of the tumour. Herein mass spectrometry provides a compelling, detailed insight into the conformational flexibility of the p53 DNA-binding domain. The plasticity of the p53 DNA-binding domain is reflected in the existence of more than one conformation, independent of any conformational changes prompted by binding. The in vacuo conformational phenotypes exhibited by common cancer-associated mutations are determined and the second-site suppressor mutation from loop L1, H115N, is probed whether it could trigger conformational changes in p53 hotspot cancer mutations. The structural basis of the binding promiscuity of p53 protein is investigated; of particular interest is the molecular interaction of the p53 N-terminus with the oncoprotein murine double minute 2, as well as with the antiapoptotic factor B-cell lymphoma-extralarge.en_US
dc.contributor.sponsorBiotechnology and Biological Sciences Research Council (BBSRC)en_US
dc.language.isoenen_US
dc.publisherThe University of Edinburghen_US
dc.relation.hasversionJurneczko, E., Cruickshank, F., Porrini, M., Clarke, D.J., Campuzano, I.D.G., Morris, M., Nikolova, P.V. and Barran, P.E. (2013) Probing the conformational diversity of cancer-associated mutations in p53 with ion-mobility mass spectrometry. Angewandte Chemie International Edition, 52, 4370-4374.en_US
dc.relation.hasversionJurneczko, E., Cruickshank, F., Porrini, M., Nikolova, P., Campuzano, I.D., Morris, M. and Barran, P.E. (2012) Intrinsic disorder in proteins: a challenge for (un)structural biology met by ion mobility-mass spectrometry. Biochemical Society Transactions, 40, 1021-1026.en_US
dc.relation.hasversionJurneczko, E., Kalapothakis, J., Campuzano, I.D., Morris, M. and Barran, P.E. (2012) Effects of drift gas on collision cross sections of a protein standard in linear drift tube and traveling wave ion mobility mass spectrometry. Analytical Chemistry, 84, 8524-8531.en_US
dc.relation.hasversionJurneczko, E. and Barran, P.E. (2011) How useful is ion mobility mass spectrometry for structural biology? The relationship between protein crystal structures and their collision cross sections in the gas phase. Analyst, 136, 20-28.en_US
dc.subjectconformationen_US
dc.subjectp53 proteinen_US
dc.subjectmass spectrometryen_US
dc.subjection mobilityen_US
dc.subjectdisordereden_US
dc.titleResolving intrinsically disordered proteins of the cancer genome with ion mobility mass spectrometryen_US
dc.typeThesis or Dissertationen_US
dc.type.qualificationlevelDoctoralen_US
dc.type.qualificationnamePhD Doctor of Philosophyen_US


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