Abstract
Compartmentalized biochemical activities are essential to all cellular processes, but there is no generalizable method to visualize dynamic protein activities in living cells at a resolution commensurate with cellular compartmentalization. here, we introduce a new class of fluorescent biosensors that detect biochemical activities in living cells at a resolution up to threefold better than the diffraction limit. these Flinc biosensors use binding-induced changes in protein fluorescence dynamics to translate kinase activities or proteinprotein interactions into changes in fluorescence fluctuations, which are quantifable through stochastic optical fluctuation imaging. A protein kinase A (PKA) biosensor allowed us to resolve minute PKA activity microdomains on the plasma membranes of living cells and to uncover the role of clustered anchoring proteins in organizing these activity microdomains. together, these fndings suggest that biochemical activities of the cell are spatially organized into an activity architecture whose structural and functional characteristics can be revealed by these new biosensors.
Original language | English (US) |
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Pages (from-to) | 427-434 |
Number of pages | 8 |
Journal | Nature Methods |
Volume | 14 |
Issue number | 4 |
DOIs | |
State | Published - Mar 13 2017 |
ASJC Scopus subject areas
- Biotechnology
- Biochemistry
- Molecular Biology
- Cell Biology