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Imaging Sites of Receptor Dephosphorylation by PTP1B on the Surface of the Endoplasmic Reticulum
Fawaz G. Haj,1Peter J. Verveer,2Anthony Squire,2Benjamin G. Neel,1*Philippe I. H. Bastiaens2*
When bound by extracellular ligands, receptor tyrosine
kinases (RTKs) on the cell surface transmit critical signals to thecell interior. Although signal termination is less well understood,protein tyrosine phosphatase-1B (PTP1B) is implicated in the
dephosphorylationand inactivation of several RTKs.
However, PTP1B resides on thecytoplasmic surface of the endoplasmic
reticulum (ER), so howand when it accesses RTKs has been
unclear. Using fluorescenceresonance energy transfer (FRET)
methods, we monitored interactionsbetween the epidermal- and
platelet-derived growth factor receptorsand PTP1B. PTP1B-catalyzed
dephosphorylation required endocytosisof the receptors and
occurred at specific sites on the surfaceof the ER. Most of the RTKs
activated at the cell surface showedinteraction with PTP1B after
internalization, establishing thatRTK activation and inactivation are
spatially and temporally partitionedwithin cells.
1 Cancer Biology Program, Division of
Hematology-Oncology, Department of Medicine, Beth Israel-Deaconess
Medical Center, Boston, MA 02115, USA.
2 European
Molecular Biology Laboratory, Meyerhofstrasse 1, 69117 Heidelberg,
Germany.
*
To whom correspondence should be addressed. E-mail:
bneel{at}caregroup.harvard.edu (B.G.N.), bastiaen{at}embl-heidelberg.de
(P.I.H.B.)
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