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A Method for the Quantitative Analysis of Protein-Protein Interactions In Vivo

dc.contributor.advisorNeumann, Heinz Prof. Dr.
dc.contributor.authorRall, Nils Arne
dc.date.accessioned2016-04-13T08:34:47Z
dc.date.available2016-04-13T08:34:47Z
dc.date.issued2016-04-13
dc.identifier.urihttp://hdl.handle.net/11858/00-1735-0000-0028-872D-1
dc.identifier.urihttp://dx.doi.org/10.53846/goediss-5609
dc.language.isodeude
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc572de
dc.titleA Method for the Quantitative Analysis of Protein-Protein Interactions In Vivoen
dc.typedoctoralThesisde
dc.contributor.refereeNeumann, Heinz Prof. Dr.
dc.date.examination2016-03-22
dc.description.abstractengThe function and activity of proteins is often modulated by other proteins they interact with. To understand cellular behaviour at the system level, a complete description of interactions among different proteins and the dynamic nature of these interactions is required.  Identification and quantification of proteins based on measured peptide intensities represents the highest standard for an unbiased and definite determination of protein interaction partners.  Inspired by the display of effectiveness of pBPA in vivo crosslinking in elucidating the mechanistic details of chromatin condensation in S. cerevisiae (Wilkins et al., 2014), the research objective of this PhD thesis was the development of a method for the quantitative identification of histone interaction partners for further decoding of chromatin structure dynamics. First, the effectiveness of pBPA in vivo crosslinking for capturing histone-protein interactions in living yeast cells was successfully confirmed. Secondly, an immunoprecipitation protocol for pulldown of HA-tagged histone pBPA mutants and their UV-induced crosslink products was established and adapted to SILAC conditions. Thirdly, initial SILAC analyses were performed and optimized until reproducible results of increased quality were obtained. Optimization led to the identification of numerous chromatin-related proteins, including a confirmed interaction partner. de
dc.contributor.coRefereeUrlaub, Henning Prof. Dr.
dc.subject.engGenetic code expansionde
dc.subject.engSILACde
dc.subject.engMass spectrometryde
dc.subject.engChromatinde
dc.subject.engHistonesde
dc.subject.engCrosslinkingde
dc.subject.engIn vivode
dc.identifier.urnurn:nbn:de:gbv:7-11858/00-1735-0000-0028-872D-1-5
dc.affiliation.instituteGöttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)de
dc.subject.gokfullBiologie (PPN619462639)de
dc.identifier.ppn857081365


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