% IMPORTANT: The following is UTF-8 encoded.  This means that in the presence
% of non-ASCII characters, it will not work with BibTeX 0.99 or older.
% Instead, you should use an up-to-date BibTeX implementation like “bibtex8” or
% “biber”.

@PHDTHESIS{Bustorff:1024924,
      author       = {Bustorff, Nuno},
      title        = {{F}olding and structural studies of saccharomyces
                      cerevisiae {P}hosphoglycerate {K}inase},
      volume       = {282},
      school       = {RWTH Aachen University},
      type         = {Dissertation},
      address      = {Jülich},
      publisher    = {Forschungszentrum Jülich GmbH Zentralbibliothek, Verlag},
      reportid     = {FZJ-2024-02574},
      isbn         = {978-3-95806-754-7},
      series       = {Schriften des Forschungszentrums Jülich Reihe
                      Schlüsseltechnologien / Key Technologies},
      pages        = {xxvi, 126},
      year         = {2024},
      note         = {Dissertation, RWTH Aachen University, 2024},
      abstract     = {Proteins are synthesized within cells by ribosomes, and
                      their functionality depend on the correct three-dimensional
                      structure obtained through the process of folding. While the
                      classical understanding of protein folding primarily focused
                      on postsynthesisfolding, recent research has shifted its
                      emphasis toward unraveling the intricacies of folding during
                      synthesis. Of particular interest in this pursuit are
                      multi-domain proteins, which constitute over $70\%$ of
                      proteins in cells. Using classical single-molecule
                      fluorescence resonance transfer efficiency (sm-FRET)
                      studies, the unfolding/refolding transitions of a two-domain
                      yeast phosphoglycerate kinase (yPGK) was explored as a model
                      for multiple domain proteins. To enhance our understanding
                      of a transition within a single-domain of full-length yPGK,
                      I assessed two FRET pair variants within the N-terminal
                      domain. Together with previous data we compared in total six
                      different variants, for which we observed three distinct
                      transitions in both domains: the first segment (positions
                      1–88 and 202–256, respectively) showed no transition
                      (i.e. no change in the distance), while the second segment
                      (positions 34–135 and 256–290, respectively) underwent a
                      conventional two-state transition. Intriguingly, labeling
                      the complete Rossmann motif (positions 1–135 and
                      202–290, respectively) revealed a compact intermediate
                      state during GuHCl-induced unfolding in transitory
                      conditions. The detailed understanding of N-terminal domain
                      transitions was essential for comparative analyses with
                      shorter-length proteins synthesized for co-translational
                      folding studies. In order to investigate the ribosomal
                      folding process of yPGK, I employed cryo-electron microscopy
                      (cryo-EM). Four distinct ribosome nascent chain complex
                      (RNC) structures were solved, each representing a nascent
                      polypeptide of a varying length. Most structures depicted
                      nascent chain density outside the ribosomal tunnel, and we
                      observed for the first time the structure of a full-length
                      protein nascent chain attached to the ribosome. In summary,
                      the presented research advances our knowledge of yPGK
                      folding transitions and offers novel routes for studying
                      co-translational folding processes within RNC complexes
                      using sm-FRET and cryo-EM.},
      cin          = {ER-C-3},
      cid          = {I:(DE-Juel1)ER-C-3-20170113},
      pnm          = {5352 - Understanding the Functionality of Soft Matter and
                      Biomolecular Systems (POF4-535) / 5241 - Molecular
                      Information Processing in Cellular Systems (POF4-524)},
      pid          = {G:(DE-HGF)POF4-5352 / G:(DE-HGF)POF4-5241},
      typ          = {PUB:(DE-HGF)3 / PUB:(DE-HGF)11},
      urn          = {urn:nbn:de:0001-20240516082247276-4907644-5},
      doi          = {10.34734/FZJ-2024-02574},
      url          = {https://juser.fz-juelich.de/record/1024924},
}