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

Rall, Nils Arne

dc.contributor.advisor	Neumann, Heinz Prof. Dr.
dc.contributor.author	Rall, Nils Arne
dc.date.accessioned	2016-04-13T08:34:47Z
dc.date.available	2016-04-13T08:34:47Z
dc.date.issued	2016-04-13
dc.identifier.uri	http://hdl.handle.net/11858/00-1735-0000-0028-872D-1
dc.identifier.uri	http://dx.doi.org/10.53846/goediss-5609
dc.language.iso	deu	de
dc.rights.uri	http://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subject.ddc	572	de
dc.title	A Method for the Quantitative Analysis of Protein-Protein Interactions In Vivo	en
dc.type	doctoralThesis	de
dc.contributor.referee	Neumann, Heinz Prof. Dr.
dc.date.examination	2016-03-22
dc.description.abstracteng	The 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.coReferee	Urlaub, Henning Prof. Dr.
dc.subject.eng	Genetic code expansion	de
dc.subject.eng	SILAC	de
dc.subject.eng	Mass spectrometry	de
dc.subject.eng	Chromatin	de
dc.subject.eng	Histones	de
dc.subject.eng	Crosslinking	de
dc.subject.eng	In vivo	de
dc.identifier.urn	urn:nbn:de:gbv:7-11858/00-1735-0000-0028-872D-1-5
dc.affiliation.institute	Göttinger Graduiertenschule für Neurowissenschaften, Biophysik und molekulare Biowissenschaften (GGNB)	de
dc.subject.gokfull	Biologie (PPN619462639)	de
dc.identifier.ppn	857081365

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Name:: Dissertation_Nils_Rall_2016_opt.pdf
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Beschreibung:: Dissertation Nils Arne Rall 2016

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Das Dokument erscheint in:

GGNB - Göttinger Graduiertenzentrum für Neurowissenschaften, Biophysik und molekulare Biowissenschaften [1115]
GGNB - Göttingen Graduate Center for Neurosciences, Biophysics and Molecular Biosciences

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