Accessible synthetic probes for staining actin inside platelets and megakaryocytes by employing Lifeact peptide
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Accessible synthetic probes for staining actin inside platelets and megakaryocytes by employing Lifeact peptide. / Cardo, Lucia; Thomas, Steven; Mazharian, Alexandra; Pikramenou, Zoe; Rappoport, Joshua; Hannon, Michael; Watson, Steve.
In: ChemBioChem, Vol. 16, No. 11, 27.07.2015, p. 1680-1688.Research output: Contribution to journal › Article › peer-review
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T1 - Accessible synthetic probes for staining actin inside platelets and megakaryocytes by employing Lifeact peptide
AU - Cardo, Lucia
AU - Thomas, Steven
AU - Mazharian, Alexandra
AU - Pikramenou, Zoe
AU - Rappoport, Joshua
AU - Hannon, Michael
AU - Watson, Steve
PY - 2015/7/27
Y1 - 2015/7/27
N2 - Lifeact is a 17-residue peptide that can be employed in cell microscopy as a probe for F-actin when fused to fluorescent proteins, but therefore is not suitable for all cell types. We have conjugated fluorescently labelled Lifeact to three different cell-penetrating systems (a myristoylated carrier (myr), the pH low insertion peptide (pHLIP) and the cationic peptide TAT) as a strategy to deliver Lifeact into cells and developed new tools for actin staining with improved synthetic accessibility and low toxicity, focusing on their suitability in platelets and megakaryocytes. Using confocal microscopy, we characterised the cell distribution of the new hybrids in fixed cells, and found that both myr- and pHLIP-Lifeact conjugates provide efficient actin staining upon cleavage of Lifeact from the carriers, without affecting cell spreading. This new approach could facilitate the design of new tools for actin visualisation.
AB - Lifeact is a 17-residue peptide that can be employed in cell microscopy as a probe for F-actin when fused to fluorescent proteins, but therefore is not suitable for all cell types. We have conjugated fluorescently labelled Lifeact to three different cell-penetrating systems (a myristoylated carrier (myr), the pH low insertion peptide (pHLIP) and the cationic peptide TAT) as a strategy to deliver Lifeact into cells and developed new tools for actin staining with improved synthetic accessibility and low toxicity, focusing on their suitability in platelets and megakaryocytes. Using confocal microscopy, we characterised the cell distribution of the new hybrids in fixed cells, and found that both myr- and pHLIP-Lifeact conjugates provide efficient actin staining upon cleavage of Lifeact from the carriers, without affecting cell spreading. This new approach could facilitate the design of new tools for actin visualisation.
KW - FRET
KW - Lifeact
KW - actin
KW - cell-penetrating hybrids
KW - fluorescent probes
U2 - 10.1002/cbic.201500120
DO - 10.1002/cbic.201500120
M3 - Article
C2 - 26062886
VL - 16
SP - 1680
EP - 1688
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 11
ER -