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@ARTICLE{Burgstaller:862520,
      author       = {Burgstaller, Sandra and Bischof, Helmut and Gensch, Thomas
                      and Stryeck, Sarah and Gottschalk, Benjamin and
                      Ramadani-Muja, Jeta and Eroglu, Emrah and Rost, Rene and
                      Balfanz, Sabine and Baumann, Arnd and Waldeck-Weiermair,
                      Markus and Hay, Jesse C. and Madl, Tobias and Graier,
                      Wolfgang F. and Malli, Roland},
      title        = {p{H}-{L}emon, a {F}luorescent {P}rotein-{B}ased p{H}
                      {R}eporter for {A}cidic {C}ompartments},
      journal      = {ACS sensors},
      volume       = {4},
      number       = {4},
      issn         = {2379-3694},
      address      = {Washington, DC},
      publisher    = {ACS Publications},
      reportid     = {FZJ-2019-02823},
      pages        = {883 - 891},
      year         = {2019},
      abstract     = {Distinct subcellular pH levels, especially in lysosomes and
                      endosomes, are essential for the degradation, modification,
                      sorting, accumulation, and secretion of macromolecules.
                      Here, we engineered a novel genetically encoded pH probe by
                      fusing the pH-stable cyan fluorescent protein (FP) variant,
                      mTurquoise2, to the highly pH-sensitive enhanced yellow
                      fluorescent protein, EYFP. This approach yielded a
                      ratiometric biosensor—referred to as pH-Lemon—optimized
                      for live imaging of distinct pH conditions within acidic
                      cellular compartments. Protonation of pH-Lemon under acidic
                      conditions significantly decreases the yellow fluorescence
                      while the cyan fluorescence increases due to reduced
                      Förster resonance energy transfer (FRET) efficiency.
                      Because of its freely reversible and ratiometric responses,
                      pH-Lemon represents a fluorescent biosensor for pH dynamics.
                      pH-Lemon also shows a sizable pH-dependent fluorescence
                      lifetime change that can be used in fluorescence lifetime
                      imaging microscopy as an alternative observation method for
                      the study of pH in acidic cellular compartments. Fusion of
                      pH-Lemon to the protein microtubule-associated protein
                      1A/1B-light chain 3B (LC3B), a specific marker of autophagic
                      membranes, resulted in its targeting within autolysosomes of
                      HeLa cells. Moreover, fusion of pH-Lemon to a
                      glycophosphatidylinositol (GPI) anchor allowed us to monitor
                      the entire luminal space of the secretory pathway and the
                      exoplasmic leaflet of the plasma membrane. Utilizing this
                      new pH probe, we revealed neutral and acidic vesicles and
                      substructures inside cells, highlighting compartments of
                      distinct pH throughout the endomembrane system. These data
                      demonstrate, that this novel pH sensor, pH-Lemon, is very
                      suitable for the study of local pH dynamics of subcellular
                      microstructures in living cells.},
      cin          = {ICS-4},
      ddc          = {570},
      cid          = {I:(DE-Juel1)ICS-4-20110106},
      pnm          = {552 - Engineering Cell Function (POF3-552)},
      pid          = {G:(DE-HGF)POF3-552},
      typ          = {PUB:(DE-HGF)16},
      pubmed       = {pmid:30864782},
      UT           = {WOS:000466442500013},
      doi          = {10.1021/acssensors.8b01599},
      url          = {https://juser.fz-juelich.de/record/862520},
}