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SHR Neuro Krebs Kardio Lipid

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 [OPEN ACCESS]
Web of Science PubMed PUBMED Central FullText FullText_MUG

 

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
Waldeck-Weiermair Markus
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Number of Figures: 6
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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
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