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@ARTICLE{Fischer:614229,
      author       = {Fischer, Alexandra A. M. and Schatz, Larissa and Baaske,
                      Julia and Römer, Winfried and Weber, Wilfried and
                      Thünauer, Roland},
      title        = {{R}eal‐time monitoring of cell surface protein arrival
                      with split luciferases},
      journal      = {Traffic},
      volume       = {24},
      number       = {10},
      issn         = {1398-9219},
      address      = {Oxford},
      publisher    = {Wiley-Blackwell},
      reportid     = {PUBDB-2024-05769},
      pages        = {453 - 462},
      year         = {2023},
      abstract     = {Each cell in a multicellular organism permanently adjusts
                      the concentration of its cell surface proteins. In
                      particular, epithelial cells tightly control the number of
                      carriers, transporters and cell adhesion proteins at their
                      plasma membrane. However, sensitively measuring the cell
                      surface concentration of a particular protein of interest in
                      live cells and in real time represents a considerable
                      challenge. Here, we introduce a novel approach based on
                      split luciferases, which uses one luciferase fragment as a
                      tag on the protein of interest and the second fragment as a
                      supplement to the extracellular medium. Once the protein of
                      interest arrives at the cell surface, the luciferase
                      fragments complement and generate luminescence. We compared
                      the performance of split Gaussia luciferase and split
                      Nanoluciferase by using a system to synchronize biosynthetic
                      trafficking with conditional aggregation domains. The best
                      results were achieved with split Nanoluciferase, for which
                      luminescence increased more than 6000-fold upon
                      recombination. Furthermore, we showed that our approach can
                      separately detect and quantify the arrival of membrane
                      proteins at the apical and basolateral plasma membrane in
                      single polarized epithelial cells by detecting the
                      luminescence signals with a microscope, thus opening novel
                      avenues for characterizing the variations in trafficking in
                      individual epithelial cells.},
      cin          = {CSSB-CF-ALFM},
      ddc          = {570},
      cid          = {I:(DE-H253)CSSB-CF-ALFM-20210629},
      pnm          = {899 - ohne Topic (POF4-899) / DFG project
                      G:(GEPRIS)390939984 - EXC 2189: CIBSS - Centre for
                      Integrative Biological Signalling Studies (390939984)},
      pid          = {G:(DE-HGF)POF4-899 / G:(GEPRIS)390939984},
      experiment   = {EXP:(DE-MLZ)NOSPEC-20140101},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:37403269},
      UT           = {WOS:001024226500001},
      doi          = {10.1111/tra.12908},
      url          = {https://bib-pubdb1.desy.de/record/614229},
}