Abstract
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.
🔬 Techniques
💻 Software
✨ Fluorophores
🧪 Sample Preparation
🔬 Cell Lines
💻 Software Details
💾 Data Repositories
🏛️ Research Organizations (ROR)
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📊 Figures
Figure 1
Isolating a subnetwork in MAP kinase signaling via genetically encoding a photocaged lysine in MEK1 active site. (a) Schematic of the MAP kinase signaling pathway and the synthetic, photoactivable sub...
Figure 2
Specific phosphorylation of ERK2 upon photoactivation of the caged MEK1. HEK293ET cells cotransfected with plasmids encoding PCKRS, pyrrolysyl tRNA CUA , EGFP-ERK2, and either A-MEK1-u0394N-HA (A, lan...
Figure 3
EGFP-ERK2 nuclear translocation upon photoactivation of the caged MEK1. (a) Representative EGFP-ERK2 subcellular fluorescence at different time points after photoactivation (2 s, 365 nm, 1 mW/cm 2 ) o...
Figure images are served from the NIH/NLM PubMed Central Open Access Subset or Europe PMC; copyright remains with the publishers and authors.
💬 Discussion
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