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J. Biol. Chem., Vol. 276, Issue 16, 12974-12982, April 20, 2001
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From § Biofrontera Pharmaceuticals AG, Hemmelratherweg
201, 51377 Leverkusen, Germany, the ** Department of Animal
Physiology, Ruhr-University of Bochum, 44780 Bochum, Germany,
By using the yeast two-hybrid system, we
previously isolated a cDNA clone encoding a novel member of the
multivalent PDZ protein family called MUPP1 containing 13 PDZ domains.
Here we report that the C terminus of the 5-hydroxytryptamine
type 2C (5-HT2C) receptor selectively interacts with
the 10th PDZ domain of MUPP1. Mutations in the extreme C-terminal SSV
sequence of the 5-HT2C receptor confirmed that the
SXV motif is critical for the interaction. Co-immunoprecipitations of MUPP1 and 5-HT2C receptors from
transfected COS-7 cells and from rat choroid plexus verified this
interaction in vivo. Immunocytochemistry revealed an
SXV motif-dependent co-clustering of both
proteins in transfected COS-7 cells as well as a colocalization in rat
choroid plexus. A 5-HT2C receptor-dependent
unmasking of a C-terminal vesicular stomatitis virus epitope of MUPP1
suggests that the interaction triggers a conformational change within
the MUPP1 protein. Moreover, 5-HT2A and 5-HT2B,
sharing the C-terminal EX(V/I)SXV sequence with
5-HT2C receptors, also bind MUPP1 PDZ domains in
vitro. The highest MUPP1 mRNA levels were
found in all cerebral cortical layers, the hippocampus, the granular
layer of the dentate gyrus, as well as the choroid plexus, where
5-HT2C receptors are highly enriched. We propose that MUPP1
may serve as a multivalent scaffold protein that selectively assembles
and targets signaling complexes.
Interaction of Serotonin 5-Hydroxytryptamine Type 2C
Receptors with PDZ10 of the Multi-PDZ Domain Protein MUPP1*
,,
,,
CNRS UPR 9023, Centre CNRS-INSERM de Pharmacologie
et Endocrinologie, 141 Rue de la Cardonille, 34094 Montpellier
Cedex 05, France, and the ¶ Department of Molecular Cell Biology,
Weizmann Institute of Science, 76100 Rehovot, Israel
*
This work was supported by grants from CNRS.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
Incumbent of the Madeleine Haas Russell Career Development Chair.
To whom correspondence should be addressed. Tel.:
49-214-8763235; Fax: 49-214-8763290; E-mail:
ullmer@biofrontera.de.
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