TY - JOUR
T1 - Live-cell imaging of enzyme-substrate interaction reveals spatial regulation of PTP1B
AU - Yudushkin, Ivan A.
AU - Schleifenbaum, Andreas
AU - Kinkhabwala, Ali
AU - Neel, Benjamin G.
AU - Schultz, Carsten
AU - Bastiaens, Philippe I.H.
PY - 2007/1/5
Y1 - 2007/1/5
N2 - Endoplasmic reticulum-localized protein-tyrosine phosphatase PTP1B terminates growth factor signal transduction by dephosphorylation of receptor tyrosine kinases (RTKs). But how PTP1B allows for RTK signaling in the cytoplasm is unclear. In order to test whether PTP1B activity is spatially regulated, we developed a method based on Förster resonant energy transfer for imaging enzyme-substrate (ES) intermediates in live cells. We observed the establishment of a steady-state ES gradient across the cell. This gradient exhibited robustness to cell-to-cell variability, growth factor activation, and RTK localization, which demonstrated spatial regulation of PTP1B activity. Such regulation may be important for generating distinct cellular environments that permit RTK signal transduction and that mediate its eventual termination.
AB - Endoplasmic reticulum-localized protein-tyrosine phosphatase PTP1B terminates growth factor signal transduction by dephosphorylation of receptor tyrosine kinases (RTKs). But how PTP1B allows for RTK signaling in the cytoplasm is unclear. In order to test whether PTP1B activity is spatially regulated, we developed a method based on Förster resonant energy transfer for imaging enzyme-substrate (ES) intermediates in live cells. We observed the establishment of a steady-state ES gradient across the cell. This gradient exhibited robustness to cell-to-cell variability, growth factor activation, and RTK localization, which demonstrated spatial regulation of PTP1B activity. Such regulation may be important for generating distinct cellular environments that permit RTK signal transduction and that mediate its eventual termination.
UR - http://www.scopus.com/inward/record.url?scp=33846678939&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=33846678939&partnerID=8YFLogxK
U2 - 10.1126/science.1134966
DO - 10.1126/science.1134966
M3 - Article
C2 - 17204654
AN - SCOPUS:33846678939
SN - 0036-8075
VL - 315
SP - 115
EP - 119
JO - Science
JF - Science
IS - 5808
ER -