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The Journal of Neuroscience, October 1, 2000, 20(19):7252-7257
PICK1 Interacts with and Regulates PKC Phosphorylation of
mGLUR7
Kumlesh K.
Dev1, 3,
Yoshiaki
Nakajima2,
Jun
Kitano1,
Steven P.
Braithwaite3,
Jeremy M.
Henley3, and
Shigetada
Nakanishi1
1 Department of Biological Sciences, Kyoto University,
Faculty of Medicine, Kyoto, 606-8501, Japan, 2 Department
of Cell Physiology, National Institute for Physiological Science,
Okazaki, 444-8585, Japan, and 3 Department of Anatomy,
University of Bristol, Medical School, Bristol, BS8 1TD, United Kingdom
The G-protein-coupled metabotropic glutamate receptor subtype 7a
(mGluR7a) is a member of group III metabotropic glutamate receptors
that plays an important role as a presynaptic receptor in regulating
transmitter release at glutamatergic synapses. Here we report that the
protein interacting with C-kinase (PICK1) binds to the C terminus (ct)
of mGluR7a. In the yeast two-hybrid system, the extreme ct of mGluR7a
was shown to interact with the PSD-95/Discs large/ZO-1 (PDZ) domain of
PICK1. Pull-down assays indicated that PICK1 was retained by a
glutathione S-transferase fusion of ct-mGluR7a. Furthermore,
recombinant and native PICK1/mGluR7a complexes were coimmunoprecipitated from COS-7 cells and rat brain tissue,
respectively. Confocal microscopy showed that both PICK1 and mGluR7a
displayed synaptic colocalization in cultured hippocampal neurons.
PICK1 has previously been shown to bind protein kinase C  -subunit
(PKC), and mGluR7a is known to be phosphorylated by PKC. We show
a relationship between these three proteins using
recombinant PICK1, mGluR7, and PKC, where they were
co-immunoprecipitated as a complex from COS-7 cells. In addition, PICK1
caused a reduction in PKC-evoked phosphorylation of mGluR7a in
in vitro phosphorylation assays. These results suggest a
role for PICK1 in modulating PKC-evoked phosphorylation of mGluR7a.
Key words:
metabotropic glutamate receptor subtype 7; protein
interacting with C-kinase (PICK1); protein kinase C; protein-protein
interactions; PDZ domain; yeast two-hybrid system; protein
phosphorylation
Copyright © 2000 Society for Neuroscience 0270-6474/00/20197252-06$05.00/0
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