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Light-Activated Kinases Enable Temporal Dissection of Signaling Networks in Living Cells

2011; American Chemical Society; Volume: 133; Issue: 7 Linguagem: Inglês

10.1021/ja1109979

1943-2984

Arnaud Gautier, Alexander Deiters, Jason W. Chin,

bioluminescence and chemiluminescence research

We report a general strategy for creating protein kinases in mammalian cells that are poised for very rapid activation by light. By photoactivating a caged version of MEK1, we demonstrate the specific, rapid, and receptor independent activation of an artificial subnetwork within the Raf/MEK/ERK pathway. Time-lapse microscopy allowed us to precisely characterize the kinetics of elementary steps in the signaling cascade and provided insight into adaptive feedback and rate-determining processes in the pathway.