%0 Journal Article %1 feldbauer2016optochemokine %A Feldbauer, Katrin %A Schlegel, Jan %A Weissbecker, Juliane %A Sauer, Frank %A Wood, Phillip G. %A Bamberg, Ernst %A Terpitz, Ulrich %D 2016 %I Public Library of Science %J PLoS ONE %K terpitz %N 10 %P e0165344-- %T Optochemokine Tandem for Light-Control of Intracellular Ca²⁺ %U http://dx.doi.org/10.1371/journal.pone.0165344 %V 11 %X <p>An optochemokine tandem was developed to control the release of calcium from endosomes into the cytosol by light and to analyze the internalization kinetics of G-protein coupled receptors (GPCRs) by electrophysiology. A previously constructed rhodopsin tandem was re-engineered to combine the light-gated Ca²⁺-permeable cation channel Channelrhodopsin-2(L132C), CatCh, with the chemokine receptor CXCR4 in a functional tandem protein tCXCR4/CatCh. The GPCR was used as a shuttle protein to displace CatCh from the plasma membrane into intracellular areas. As shown by patch-clamp measurements and confocal laser scanning microscopy, heterologously expressed tCXCR4/CatCh was internalized via the endocytic SDF1/CXCR4 signaling pathway. The kinetics of internalization could be followed electrophysiologically via the amplitude of the CatCh signal. The light-induced release of Ca²⁺ by tandem endosomes into the cytosol via CatCh was visualized using the Ca²⁺-sensitive dyes rhod2 and rhod2-AM showing an increase of intracellular Ca²⁺ in response to light.</p>

@article{feldbauer2016optochemokine, abstract = {<p>An optochemokine tandem was developed to control the release of calcium from endosomes into the cytosol by light and to analyze the internalization kinetics of G-protein coupled receptors (GPCRs) by electrophysiology. A previously constructed rhodopsin tandem was re-engineered to combine the light-gated Ca²⁺-permeable cation channel Channelrhodopsin-2(L132C), CatCh, with the chemokine receptor CXCR4 in a functional tandem protein tCXCR4/CatCh. The GPCR was used as a shuttle protein to displace CatCh from the plasma membrane into intracellular areas. As shown by patch-clamp measurements and confocal laser scanning microscopy, heterologously expressed tCXCR4/CatCh was internalized via the endocytic SDF1/CXCR4 signaling pathway. The kinetics of internalization could be followed electrophysiologically via the amplitude of the CatCh signal. The light-induced release of Ca²⁺ by tandem endosomes into the cytosol via CatCh was visualized using the Ca²⁺-sensitive dyes rhod2 and rhod2-AM showing an increase of intracellular Ca²⁺ in response to light.</p>}, added-at = {2016-10-24T08:43:29.000+0200}, author = {Feldbauer, Katrin and Schlegel, Jan and Weissbecker, Juliane and Sauer, Frank and Wood, Phillip G. and Bamberg, Ernst and Terpitz, Ulrich}, biburl = {https://www.bibsonomy.org/bibtex/22c54a2193e26f4afc4d69b784e0ed369/reichert}, interhash = {0c7ad2472d239915dc99b7223e79d414}, intrahash = {2c54a2193e26f4afc4d69b784e0ed369}, journal = {PLoS ONE}, keywords = {terpitz}, month = oct, number = 10, pages = {e0165344--}, publisher = {Public Library of Science}, timestamp = {2016-10-24T09:09:57.000+0200}, title = {Optochemokine Tandem for Light-Control of Intracellular Ca²⁺}, url = {http://dx.doi.org/10.1371/journal.pone.0165344}, volume = 11, year = 2016 }