NIDD, a Novel DHHC-containing Protein, Targets Neuronal Nitric-oxide Synthase (nNOS) to the Synaptic Membrane through a PDZ-dependent Interaction and Regulates nNOS Activity
Targeting of neuronal nitric-oxide synthase (nNOS) to appropriate sites in a cell is mediated by interactions with its PDZ domain and plays an important role in specifying the sites of reaction of nitric oxide (NO) in the central nervous system. Here we report the identification and characterization...
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Published in | The Journal of biological chemistry Vol. 279; no. 28; pp. 29461 - 29468 |
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Main Authors | , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
American Society for Biochemistry and Molecular Biology
09.07.2004
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Subjects | |
Online Access | Get full text |
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Summary: | Targeting of neuronal nitric-oxide synthase (nNOS) to appropriate sites in a cell is mediated by interactions with its PDZ
domain and plays an important role in specifying the sites of reaction of nitric oxide (NO) in the central nervous system.
Here we report the identification and characterization of a novel n NOS- i nteracting D HHC d omain-containing protein with d endritic mRNA (NIDD) (GenBank⢠accession number AB098078), which increases nNOS enzyme activity by targeting the nNOS to the
synaptic plasma membrane in a PDZ domain-dependent manner. The deduced NIDD protein consisted of 392 amino acid residues and
possessed five transmembrane segments, a zinc finger DHHC domain, and a PDZ-binding motif (-EDIV) at its C-terminal tail.
In vitro pull-down assays suggested that the C-terminal tail region of NIDD specifically interacted with the PDZ domain of nNOS. The
PDZ dependence was confirmed by an experiment using a deletion mutant, and the interaction was further confirmed by co-sedimentation
assays using COS-7 cells transfected with NIDD and nNOS. Both NIDD and nNOS were enriched in synaptosome and synaptic plasma
membrane fractions and were present in the lipid raft and postsynaptic density fractions in the rat brain. Co-localization
of these proteins was also observed by double staining of the proteins in cultured cortical neurons. Thus, NIDD and nNOS were
co-localized in the brain, although the colocalizing regions were restricted, as indicated by the distribution of their mRNA
expression. Most important, co-transfection of NIDD and nNOS increased NO-producing nNOS activity. These results suggested
that NIDD plays an important role in the regulation of the NO signaling pathway at postsynaptic sites through targeting of
nNOS to the postsynaptic membrane. |
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Bibliography: | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 |
ISSN: | 0021-9258 1083-351X |
DOI: | 10.1074/jbc.M401471200 |