Format

Send to

Choose Destination
See comment in PubMed Commons below
Cell. 2014 Jun 19;157(7):1724-34. doi: 10.1016/j.cell.2014.04.039.

High-sensitivity measurements of multiple kinase activities in live single cells.

Author information

1
Department of Bioengineering, Stanford University, Stanford, CA 94305, USA. Electronic address: sregot@stanford.edu.
2
Department of Bioengineering, Stanford University, Stanford, CA 94305, USA.
3
Department of Bioengineering, Stanford University, Stanford, CA 94305, USA. Electronic address: mcovert@stanford.edu.

Abstract

Increasing evidence has shown that population dynamics are qualitatively different from single-cell behaviors. Reporters to probe dynamic, single-cell behaviors are desirable yet relatively scarce. Here, we describe an easy-to-implement and generalizable technology to generate reporters of kinase activity for individual cells. Our technology converts phosphorylation into a nucleocytoplasmic shuttling event that can be measured by epifluorescence microscopy. Our reporters reproduce kinase activity for multiple types of kinases and allow for calculation of active kinase concentrations via a mathematical model. Using this technology, we made several experimental observations that had previously been technicallyunfeasible, including stimulus-dependent patterns of c-Jun N-terminal kinase (JNK) and nuclear factor kappa B (NF-κB) activation. We also measured JNK, p38, and ERK activities simultaneously, finding that p38 regulates the peak number, but not the intensity, of ERK fluctuations. Our approach opens the possibility of analyzing a wide range of kinase-mediated processes in individual cells.

Comment in

PMID:
24949979
PMCID:
PMC4097317
DOI:
10.1016/j.cell.2014.04.039
[Indexed for MEDLINE]
Free PMC Article
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Elsevier Science Icon for PubMed Central
    Loading ...
    Support Center