The neuronal nitric oxide synthase PDZ motif binds to ‐G(D,E)XV<sup>*</sup> carboxyterminal sequences

Schepens, Jan; Cuppen, Edwin; Wieringa, B.; Hendriks, Wiljan

doi:10.1016/s0014-5793(97)00481-x

Cited by 53 publications

(58 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Therefore we wanted to determine whether there was a specific interaction between fulllength a 1A -ARs and nNOS in intact cells. These results support an interaction between nNOS and a 1A -ARs, which could be due to the previously reported interaction of the receptor C-terminus and the PDZ domain of nNOS [2]. However, this hypothesis is weakened by the unexpected observation that nNOS also co-immunoprecipitates with both a 1B and a 1D -ARs.…”

Section: Discussionsupporting

confidence: 88%

“…Previous work using a yeast two hybrid assay showed that the C-terminal 114 amino acids of rat a 1A -ARs (referred to by the previous name of a 1C -) strongly interacted with residues 1-111 of nNOS [2]. The bradykinin B2 receptor, also a G protein coupled receptor, has been shown to bind directly to the oxygenase domain of nNOS and form an inhibitory complex [8], and it has been proposed that nNOS is released and activated upon receptor stimulation.…”

Section: Discussionmentioning

confidence: 99%

“…Experiments using the yeast two-hybrid system showed previously that a protein corresponding to the last 114 amino acids of the rat a 1A -AR (previously referred to as a 1C -AR) interacted strongly with the PDZ domain of nNOS [2]. Since the corresponding amino acids at the C-terminus of a 1B (PGQF) and a 1D -ARs (ETDI) would not be predicted to interact with this PDZ domain, an interaction between a 1A -ARs and nNOS could represent an interaction unique to this subtype.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Interaction of neuronal nitric oxide synthase with alpha1-adrenergic receptor subtypes in transfected HEK-293 cells

Pupo

Minneman

2002

BMC Pharmacol

View full text Add to dashboard Cite

show abstract

Section: Discussionsupporting

confidence: 88%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Interaction of neuronal nitric oxide synthase with alpha1-adrenergic receptor subtypes in transfected HEK-293 cells

Pupo

Minneman

2002

BMC Pharmacol

View full text Add to dashboard Cite

show abstract

“…nNOS Binds CtBP at Its Peptide-binding Site-It has been shown previously that the PDZ domain of nNOS preferentially binds to peptides ending with the sequence G(D/E)AV (22,23) or EXAV (15,16). The last four amino acids of CtBP are SDQL, where the D fits the consensus for the second amino acid from the carboxyl terminus, and the L is a conservative substitution for the carboxyl-terminal V. To determine whether CtBP binds to the peptide-binding site in the PDZ domain of nNOS, we performed affinity chromatography of Cos-7 cells transfected with T7-tagged CtBP using a GST-nNOS-PDZ column.…”

Section: Isolation Of a 48-kda Protein That Binds To The Pdz Domain Omentioning

confidence: 99%

Binding of Neuronal Nitric-oxide Synthase (nNOS) to Carboxyl-terminal-binding Protein (CtBP) Changes the Localization of CtBP from the Nucleus to the Cytosol

Riefler

Firestein

2001

Journal of Biological Chemistry

View full text Add to dashboard Cite

Recent work suggests a role for PDZ domains in the targeting of binding partners to specific sites in the cell. To identify whether the PDZ domain of neuronal nitricoxide synthase (nNOS) can play such a role, we performed affinity chromatography of brain extract with the nNOS PDZ domain. We identified the carboxyl-terminal-binding protein (CtBP), a phosphoprotein first identified as a binding partner to adenovirus E1A, as a nNOS binding partner. CtBP interacts with the PDZ domain of nNOS, and this interaction can be competed with peptide that binds to the PDZ peptide-binding site. In addition, binding of CtBP to nNOS is dependent on its carboxyl-terminal sequence -DXL, residues conserved between species that fit the canonical sequence for nNOS PDZ binding. Immunoprecipitation studies show that CtBP and nNOS associate in the brain. When CtBP is expressed in Madin-Darby canine kidney cells, its distribution is primarily nuclear; however, when CtBP is co-expressed with nNOS, its localization becomes more cytosolic. This change in CtBP localization does not occur when its carboxyl-terminal nNOS PDZ binding motif is mutated or when CtBP is co-expressed with postsynaptic density 95, another PDZ domain-containing protein. Taken together, our data suggest a new function for nNOS as a regulator of CtBP nuclear localization. Nitric oxide (NO)1 is a gaseous molecule that plays an important role in the central nervous system (1, 2). It is produced by the enzyme neuronal nitric-oxide synthase (nNOS), which can be stimulated when glutamate receptors of the N-methyl-D-aspartate receptor family are activated, and calcium enters the neuron (2). Calcium binds to calmodulin, and the complex then activates nNOS. Furthermore, the NO produced can increase the production of 3Ј,5Ј-cyclic guanosine monophosphate levels, which can activate other enzymes including protein kinases (3, 4). Recent evidence supports a role for NO in synaptic plasticity (5), long-term potentiation (6), and aspects of learning and memory (7).nNOS contains a PDZ (PSD-95, discs-large and zona occludens-1) domain, a consensus sequence of approximately 90 amino acids that has been shown to mediate protein-protein interactions (8). In neurons, nNOS is targeted to synaptic sites via its interaction with the PDZ domains of PSD-95 and PSD-93 (9). In fact, PSD-95 also interacts with N-methyl-Daspartate receptors via one of its PDZ domains. Through its concurrent interaction with nNOS, PSD-95 serves as a physical tether to allow nNOS signaling by N-methyl-D-aspartate receptor activity (10, 11). By abolishing expression of PSD-95 protein either by knockout technology in mice or by antisense technology in tissue culture, it has been shown that the presence of PSD-95 is essential for NO production by glutamate stimulation (12, 13). Thus, the PDZ domain of nNOS plays an important role in helping to localize nNOS to appropriate sites in the neuron.Subcellular fractionation shows that roughly half of nNOS is soluble, and half is particulate (14). As such, binding partners of t...

show abstract

“…PDZ domains are globular structures that function as specific protein-recognition modules in a variety of cellular processes including the binding of carboxyterminus of E6 proteins (Harris and Lim, 2001;Thomas et al, 2001;Lee et al, 1997). In fact, a computer search program (http://scansite.mit.edu; Yaffe et al, 2001) identified the sequence, 35 GKGTSAADAVEVPAP 48 which immediately follows the L2G box in MGMT, as a class 3 PDZ site, originally identified in the neuronal nitric oxide synthase (nNOS) (Schepens et al, 1997). These structural considerations lend support to our observations that MGMT is a target for the E6/E6-AP system, however, the actual involvement of these motifs remains to be confirmed through mutation studies.…”

mentioning

confidence: 99%

The DNA repair protein, O6-Methylguanine-DNA methyltransferase is a proteolytic target for the E6 human papillomavirus oncoprotein

Srivenugopal

Ali‐Osman

2002

Oncogene

View full text Add to dashboard Cite

We have previously shown that O 6 -methylguanine-DNA methyltransferase (MGMT), a DNA repair protein that protects tissues against toxic and carcinogenic effects of alkylating agents, is degraded through ubiquitinationdependent proteolysis. Here, we investigated the role of the human papillomavirus (HPV) E6 protein in MGMT degradation. In three pairs of isogenic human tumor cell lines in which a member of each pair expressed the E6 protein through stable transfection (HCT116/HCT116-E6, MCF7/MCF7-E6, and RKO/RKO-E6), we found a consistent 40 -55% reduction in the MGMT protein level and its activity in all E6-expressing cells compared with the parent cells (P=50.05). E6 expression did not, however, alter the levels of MGMT mRNA. Addition of the recombinant MGMT (rMGMT) protein to extracts of HCT116/E6 cells resulted in the binding of E6 to MGMT. Further, the purified E6 protein promoted the degradation of rMGMT in rabbit reticulocyte lysates. Immunoprecipitation assays showed the presence of a ternary protein complex between MGMT, E6, and the cellular ubiquitin-ligase E6-associated protein (E6-AP). Transient transfection of the p53-null H1299 lung tumor cells with an E6 construct also down-regulated the MGMT. The MGMT protein also showed structural features that are compatible for interaction with the E6, and E6-AP components. Collectively, these data suggest that the oncogenic E6 proteins enhance the ubiquitindependent proteolysis of MGMT.

show abstract

The neuronal nitric oxide synthase PDZ motif binds to ‐G(D,E)XV^* carboxyterminal sequences

Cited by 53 publications

References 38 publications

Interaction of neuronal nitric oxide synthase with alpha1-adrenergic receptor subtypes in transfected HEK-293 cells

Interaction of neuronal nitric oxide synthase with alpha1-adrenergic receptor subtypes in transfected HEK-293 cells

Binding of Neuronal Nitric-oxide Synthase (nNOS) to Carboxyl-terminal-binding Protein (CtBP) Changes the Localization of CtBP from the Nucleus to the Cytosol

The DNA repair protein, O6-Methylguanine-DNA methyltransferase is a proteolytic target for the E6 human papillomavirus oncoprotein

Contact Info

Product

Resources

About

The neuronal nitric oxide synthase PDZ motif binds to ‐G(D,E)XV* carboxyterminal sequences

Cited by 53 publications

References 38 publications

Interaction of neuronal nitric oxide synthase with alpha1-adrenergic receptor subtypes in transfected HEK-293 cells

Interaction of neuronal nitric oxide synthase with alpha1-adrenergic receptor subtypes in transfected HEK-293 cells

Binding of Neuronal Nitric-oxide Synthase (nNOS) to Carboxyl-terminal-binding Protein (CtBP) Changes the Localization of CtBP from the Nucleus to the Cytosol

The DNA repair protein, O6-Methylguanine-DNA methyltransferase is a proteolytic target for the E6 human papillomavirus oncoprotein

Contact Info

Product

Resources

About

The neuronal nitric oxide synthase PDZ motif binds to ‐G(D,E)XV^* carboxyterminal sequences