TY - JOUR
T1 - Temporal Analysis of PP2A Phosphatase Activity During Insulin Stimulation Using a Direct Activity Probe
AU - Beck, Jon R.
AU - Truong, Tiffany
AU - Stains, Cliff I.
N1 - Funding Information:
We thank the Nebraska Center for Mass Spectrometry for assistance with characterization of peptide substrates and small molecules. Funding was provided by the proposed Center for Integrated Biomolecular Communication, the University of NebraskañYLincoln, and the National Institutes of Health (R35GM119751).
Publisher Copyright:
© 2016 American Chemical Society.
PY - 2016/12/16
Y1 - 2016/12/16
N2 - Protein serine/threonine phosphatases (PSPs) are ubiquitously expressed in mammalian cells. In particular, PP2A accounts for up to 1% of the total protein within cells. Despite clear evidence for the role of PP2A in cellular signaling, there is a lack of information concerning the magnitude and temporal dynamics of PP2A catalytic activity during insulin stimulation. Herein, we describe the development of a direct, fluorescent activity probe capable of reporting on global changes in PP2A enzymatic activity in unfractionated cell lysates. Utilizing this new probe, we profiled the magnitude as well as temporal dynamics of PP2A activity during insulin stimulation of liver hepatocytes. These results provide direct evidence for the rapid response of PP2A catalytic activity to extracellular stimulation, as well as insight into the complex regulation of phosphorylation levels by opposing kinase and phosphatase activities within the cell. This study provides a new tool for investigating the chemical biology of PSPs.
AB - Protein serine/threonine phosphatases (PSPs) are ubiquitously expressed in mammalian cells. In particular, PP2A accounts for up to 1% of the total protein within cells. Despite clear evidence for the role of PP2A in cellular signaling, there is a lack of information concerning the magnitude and temporal dynamics of PP2A catalytic activity during insulin stimulation. Herein, we describe the development of a direct, fluorescent activity probe capable of reporting on global changes in PP2A enzymatic activity in unfractionated cell lysates. Utilizing this new probe, we profiled the magnitude as well as temporal dynamics of PP2A activity during insulin stimulation of liver hepatocytes. These results provide direct evidence for the rapid response of PP2A catalytic activity to extracellular stimulation, as well as insight into the complex regulation of phosphorylation levels by opposing kinase and phosphatase activities within the cell. This study provides a new tool for investigating the chemical biology of PSPs.
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U2 - 10.1021/acschembio.6b00697
DO - 10.1021/acschembio.6b00697
M3 - Article
C2 - 27805358
AN - SCOPUS:85006354888
SN - 1554-8929
VL - 11
SP - 3284
EP - 3288
JO - ACS chemical biology
JF - ACS chemical biology
IS - 12
ER -