Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
SHR Neuro Krebs Kardio Lipid Stoffw Microb
Waldeck-Weiermair, M; Bischof, H; Blass, S; Deak, AT; Klec, C; Graier, T; Roller, C; Rost, R; Eroglu, E; Gottschalk, B; Hofmann, NA; Graier, WF; Malli, R.
Generation of Red-Shifted Cameleons for Imaging Ca²⁺ Dynamics of the Endoplasmic Reticulum.
Sensors (Basel). 2015; 15(6):13052-13068
Doi: 10.3390/s150613052
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- Führende Autor*innen der Med Uni Graz
- Waldeck-Weiermair Markus
- Co-Autor*innen der Med Uni Graz
- Bischof Helmut
- Blass Sandra
- Deak Andras Tamas
- EROGLU Emrah
- Gottschalk Benjamin
- Graier Thomas
- Graier Wolfgang
- Hofmann Nicole
- Klec Christiane
- Malli Roland
- Rost René
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- Abstract:
- Cameleons are sophisticated genetically encoded fluorescent probes that allow quantifying cellular Ca2+ signals. The probes are based on Förster resonance energy transfer (FRET) between terminally located fluorescent proteins (FPs), which move together upon binding of Ca2+ to the central calmodulin myosin light chain kinase M13 domain. Most of the available cameleons consist of cyan and yellow FPs (CFP and YFP) as the FRET pair. However, red-shifted versions with green and orange or red FPs (GFP, OFP, RFP) have some advantages such as less phototoxicity and minimal spectral overlay with autofluorescence of cells and fura-2, a prominent chemical Ca2+ indicator. While GFP/OFP- or GFP/RFP-based cameleons have been successfully used to study cytosolic and mitochondrial Ca2+ signals, red-shifted cameleons to visualize Ca2+ dynamics of the endoplasmic reticulum (ER) have not been developed so far. In this study, we generated and tested several ER targeted red-shifted cameleons. Our results show that GFP/OFP-based cameleons due to miss-targeting and their high Ca2+ binding affinity are inappropriate to record ER Ca2+ signals. However, ER targeted GFP/RFP-based probes were suitable to sense ER Ca2+ in a reliable manner. With this study we increased the palette of cameleons for visualizing Ca2+ dynamics within the main intracellular Ca2+ store.
- Find related publications in this database (using NLM MeSH Indexing)
- Calcium - analysis
- Endoplasmic Reticulum - chemistry
- Fluorescence Resonance Energy Transfer -
- Fluorescent Dyes - chemistry
- HEK293 Cells -
- HeLa Cells -
- Humans -
- Luminescent Proteins - chemistry
- Microscopy, Confocal -
- Find related publications in this database (Keywords)
- Forster resonance energy transfer
- FRET
- calcium
- endoplasmic reticulum
- store operated calcium entry
- SOCE
- clover
- mRuby2
- dissociation constant