2017
Mo, Gary C H; Ross, Brian; Hertel, Fabian; Manna, Premashis; Yang, Xinxing; Greenwald, Eric; Booth, Chris; Plummer, Ashlee M; Tenner, Brian; Chen, Zan; Wang, Yuxiao; Kennedy, Eileen J; Cole, Philip A; Fleming, Karen G; Palmer, Amy; Jimenez, Ralph; Xiao, Jie; Dedecker, Peter; Zhang, Jin
Genetically encoded biosensors for visualizing live-cell biochemical activity at super-resolution Journal Article
In: Nat Methods, vol. 14, no. 4, pp. 427-434, 2017, ISSN: 1548-7105.
Abstract | Links | BibTeX | Tags: Biosensing Techniques, Cell Membrane, Cyclic AMP-Dependent Protein Kinases, Escherichia coli, Fluorescence Resonance Energy Transfer, Fluorescent Dyes, Green Fluorescent Proteins, HeLa Cells, Humans, Microscopy, Molecular Imaging, Mutagenesis, Protein Interaction Mapping, Site-Directed, Stochastic Processes
@article{1299945,
title = {Genetically encoded biosensors for visualizing live-cell biochemical activity at super-resolution},
author = {Gary C H Mo and Brian Ross and Fabian Hertel and Premashis Manna and Xinxing Yang and Eric Greenwald and Chris Booth and Ashlee M Plummer and Brian Tenner and Zan Chen and Yuxiao Wang and Eileen J Kennedy and Philip A Cole and Karen G Fleming and Amy Palmer and Ralph Jimenez and Jie Xiao and Peter Dedecker and Jin Zhang},
doi = {10.1038/nmeth.4221},
issn = {1548-7105},
year = {2017},
date = {2017-04-01},
journal = {Nat Methods},
volume = {14},
number = {4},
pages = {427-434},
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 protein-protein interactions into changes in fluorescence fluctuations, which are quantifiable 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 findings 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.},
keywords = {Biosensing Techniques, Cell Membrane, Cyclic AMP-Dependent Protein Kinases, Escherichia coli, Fluorescence Resonance Energy Transfer, Fluorescent Dyes, Green Fluorescent Proteins, HeLa Cells, Humans, Microscopy, Molecular Imaging, Mutagenesis, Protein Interaction Mapping, Site-Directed, Stochastic Processes},
pubstate = {published},
tppubtype = {article}
}