Genetically encoded FRET probe for PKC activity based on pleckstrin

Journal of the American Chemical Society
Andreas SchleifenbaumCarsten Schultz

Abstract

We developed a probe for investigating protein kinase C (PKC) activity in living cells. The probe is based on a fragment of pleckstrin enclosed by two FRET-capable fluorophores. PKC activity modulation was reliably followed by FRET change in vitro and in vivo. The probe responds quickly to PKC activation by phorbol ester. FRET changes were reversible when PKC inhibitors were administered. Stimulation of cellular signaling pathways using histamine or bradykinin triggered PKC in a physiologically relevant way.

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Citations

Jan 16, 2013·Journal of the American Chemical Society·Peter DedeckerJohan Hofkens
Mar 23, 2005·Nature Methods·Melissa D ShultsBarbara Imperiali
Apr 5, 2007·Nature Protocols·Justin BrumbaughCarsten Schultz
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Jun 2, 2011·Cold Spring Harbor Perspectives in Biology·Peter Lipp, Gregor Reither
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Nov 1, 2007·HFSP Journal·Carsten Schultz
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