CDC STACKS serves as an archival repository of CDC-published products including scientific findings, journal articles, guidelines, recommendations, or other public health information authored or co-authored by CDC or funded partners.
As a repository, CDC STACKS retains documents in their original published format to ensure public access to scientific information.
i
High-sensitivity measurements of multiple kinase activities in live single cells
-
Jun 19 2014
-
-
Source: Cell. 2014; 157(7):1724-1734.
Details:
-
Alternative Title:Cell
-
Personal Author:
-
Description: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.
-
Subjects:
-
Source:
-
Pubmed ID:24949979
-
Pubmed Central ID:PMC4097317
-
Document Type:
-
Funding:
-
Volume:157
-
Issue:7
-
Collection(s):
-
Main Document Checksum:
-
Download URL:
-
File Type: