Imaging kinase activity at protein scaffolds.
Kinase signaling is under tight spatiotemporal control, with signaling hubs within the cell often coordinated by protein scaffolds. Genetically encoded kinase activity reporters afford a unique tool to interrogate the rate, amplitude, and duration of kinase signaling at specific locations throughout the cell. This protocol describes how to assay kinase activity at a protein scaffold in live cells using a fluorescence resonance energy transfer (FRET)-based kinase activity sensor for protein kinase D (PKD) as an example.
Department of Pharmacology, University of California at San Diego, San Diego, CA, USA.
Maya T
MT
Kunkel
Alexandra C
AC
Newton
D000818:Animals; D002470:Cell Survival; D004285:Dogs; D057075:Enzyme Assays / Q000379:methods*; D031541:Fluorescence Resonance Energy Transfer; D061985:Madin Darby Canine Kidney Cells; D057054:Molecular Imaging / Q000379:methods*; D011494:Protein Kinases / Q000737:chemistry / Q000378:metabolism*; D017434:Protein Structure, Tertiary
D011494:Protein Kinases