Optochemokine tandem for light-control of intracellular Ca2+

  • 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.
Author:Katrin Feldbauer, Jan Schlegel, Juliane WeissbeckerGND, Frank Sauer, Phillip G. Wood, Ernst BambergGND, Ulrich Terpitz
Pubmed Id:https://pubmed.ncbi.nlm.nih.gov/27768773
Parent Title (English):PLoS one
Place of publication:Lawrence, Kan.
Contributor(s):Alexander G. Obukhov
Document Type:Article
Date of Publication (online):2016/10/21
Date of first Publication:2016/10/21
Publishing Institution:Universitätsbibliothek Johann Christian Senckenberg
Release Date:2016/11/10
Issue:(10): e0165344
Page Number:20
First Page:1
Last Page:20
Copyright: © 2016 Feldbauer et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Institutes:Biochemie, Chemie und Pharmazie / Biochemie und Chemie
Angeschlossene und kooperierende Institutionen / MPI für Biophysik
Dewey Decimal Classification:5 Naturwissenschaften und Mathematik / 57 Biowissenschaften; Biologie / 570 Biowissenschaften; Biologie
Licence (German):License LogoCreative Commons - Namensnennung 4.0