Characterization of a 7.5-kDa Protein Kinase C Substrate (RC3 Protein, Neurogranin) from Rat Brain

Huang, Kuo‐Ping; Huang, Freesia L.; Chen, Hao-Chia

doi:10.1006/abbi.1993.1463

Cited by 91 publications

(100 citation statements)

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“…The phosphorylated Ng also stimulates the G-protein-coupled phosphoinositide second messenger pathways that trigger the mobilization of Ca 2ϩ from intracellular stores (8). PKC is the only known Ng kinase in vitro (5), and deletion of PKC ␥ gene in mice negates both the glutamate-and depolarization-mediated phosphorylation of Ng (9). The close functional relationship between Ng and PKC ␥ is further illustrated by their similar developmental expression patterns in the cerebral cortex (10)(11)(12).…”

mentioning

confidence: 88%

Involvement of neurogranin in the modulation of calcium/calmodulin-dependent protein kinase II, synaptic plasticity, and spatial learning: A study with knockout mice

Pak

Huang

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

231

229

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Neurogranin͞RC3 is a neural-specific Ca 2؉ -sensitive calmodulin (CaM)-binding protein whose CaM-binding affinity is modulated by phosphorylation and oxidation. Here we show that deletion of the Ng gene in mice did not result in obvious developmental or neuroanatomical abnormalities but caused an impairment of spatial learning and changes in hippocampal short-and long-term plasticity (paired-pulse depression, synaptic fatigue, long-term potentiation induction). These deficits were accompanied by a decreased basal level of the activated Ca 2؉ ͞CaM-dependent kinase II (CaMKII) (Ϸ60% of wild type). Furthermore, hippocampal slices of the mutant mice displayed a reduced ability to generate activated CaMKII after stimulation of protein phosphorylation and oxidation by treatments with okadaic acid and sodium nitroprusside, respectively. These results indicate a central role of Ng in the regulation of CaMKII activity with decisive influences on synaptic plasticity and spatial learning.

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mentioning

confidence: 88%

Involvement of neurogranin in the modulation of calcium/calmodulin-dependent protein kinase II, synaptic plasticity, and spatial learning: A study with knockout mice

Pak

Huang

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

231

229

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“…The differences in the radioactivities among the various protein bands from those samples treated with or without 50 mM DTT reflected the extent of protein S-thiolation. The same membrane was used for the identification of Nm and Ng by immunoblot with a preparation of antibody (number 270) recognizing both proteins (34).…”

Section: Characterization Of Protein S-thiolation-[mentioning

confidence: 99%

“…For analysis of the kinetic parameters of GS-Ng and GS-Nm phosphorylations by PKC, DTT was omitted. 32 P incorporations into Ng and Nm were measured by the Dowex AG1 ϫ8/DEAE-cellulose mini column method (34). Kinetic constants were determined using the ALLFIT program (37).…”

Section: Characterization Of Protein S-thiolation-[mentioning

confidence: 99%

“…Preparation of Rat Brain Ng and Nm and Recombinant Ng-Rat brain Ng (34) and Nm (35) and the recombinant Ng (36) were purified to homogeneity as described previously. For the preparations of reduced Ng (red-Ng) and Nm (red-Nm), purified proteins were incubated in 20 mM Tris-Cl, pH 7.5, 1 mM EDTA, and 1 mM DTT for 10 min.…”

Section: Characterization Of Protein S-thiolation-[mentioning

confidence: 99%

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Glutathiolation of Proteins by Glutathione DisulfideS-Oxide Derived from S-Nitrosoglutathione

Huang

2001

Journal of Biological Chemistry

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S-Nitrosoglutathione (GSNO) undergoes spontaneous degradation that generates several nitrogen-containing compounds and oxidized glutathione derivatives. We identified glutathione sulfonic acid, glutathione disulfide S-oxide (GS(O)SG), glutathione disulfide S-dioxide, and GSSG as the major decomposition products of GSNO. Each of these compounds and GSNO were tested for their efficacies to modify rat brain neurogranin/RC3 (Ng) and neuromodulin/GAP-43 (Nm). Among them, GS(O)SG was found to be the most potent in causing glutathiolation of both proteins; four glutathiones were incorporated into the four Cys residues of Ng, and two were incorporated into the two Cys residues of Nm. Protein S-glutathiolation can be induced in cells by mild oxidative stress (1). GSSG has been shown to oxidatively regulate the activity of several purified enzymes including carbonic anhydrase III (2, 3), protein kinase C (PKC) 1 (4), human aldose reductase (5), and human immunodeficiency virus, type I protease (6), and in each case the effects of glutathiolation can be reversed by reducing agents. As the concentration of reduced GSH in the mammalian cells is in the millimolar range and that of GSSG is less than 5% of GSH, glutathiolation of proteins by GSSG in vivo is not likely an efficient mechanism. More recently, the superoxide-induced glutathiolation of protein (7) and that induced by peroxynitrite, nitric oxide (NO), and nitrosothiol, in particular, S-nitrosoglutathione (GSNO), are thought to be the main avenues leading to protein Sthiolation (8 -12). In mammalian cells, a relatively high concentration of GSH (0.5-10 mM) serves as an NO sink to form GSNO (13-16), which can undergo transnitrosylation with protein sulfhydryl group to form S-nitrosoprotein and GSH or to form protein-GSH mixed disulfide and nitroxyl (17)(18)(19)(20). GSNO can also release NO in the presence of cuprous ion (21), ascorbate (22), or thiols (20, 23) and serves as a possible source of nitrsonium or nitroxyl ions (24). In addition, GSNO is unstable in aqueous solution and undergoes decomposition, which is believed to be homolytic cleavage of the S-N bond to give NO and a thiyl radical (25,26). Indeed, the reactions involving GSNO are fairly complex and generate many potential products including ammonia, NO, nitrous oxide, nitrite, sulfinamide, hydroxylamine, and several oxidized forms of glutathione (20, 23). Recently, it was found that freshly prepared GSNO was effective in S-nitrosylation of proteins through transnitrosylation, whereas the decomposed GSNO was more effective in S-glutathiolation of proteins (10). It was suggested that glutathione sulfenic acid was the active component for glutathiolation of proteins.Neurogranin/RC3 (Ng) and neuromodulin/GAP-43 (Nm) are two prominent PKC substrates in the brain. Phoshorylations of both Ng and Nm reduce their binding affinities for calmodulin (CaM) (27,28). Rat brain Ng contains four, and Nm contains two Cys residues; these Cys residues in Ng form two pairs of intramolecular disulfides upon oxidation by NO and...

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“…This postsynaptic PKC substrate, purified to homogeneity from bovine forebrain, consists of 78 amino acid residues, has a molecular mass of 7.8 kDa and is a member of the family of acidic substrates of PKC (pI 5.6) [12,38].…”

Section: Neurograninmentioning

confidence: 99%

Long-term potentiation and synaptic protein phosphorylation

Pasinelli

Ramakers

Urban

et al. 1995

Behavioural Brain Research

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Long-term potentiation (LTP) is a well known experimental model for studying the activity-dependent enhancement of synaptic plasticity, and because of its long duration and its associative properties, it has been proposed as a system to investigate the molecular mechanisms of memory formation. At present, there are several lines of evidence that indicate that pre-and postsynaptic kinases and their specific substrates are involved in molecular mechanisms underlying LTP. Many studies focus on the involvement of protein kinase C (PKC). One way to investigate the role of PKC in long-term potentiation is to determine the degree of phosphorylation of its substrates after in situ phosphorylation in hippocampal slices. Two possible targets are the presynaptic membrane-associated protein B-50 (a.k.a. GAP 43, neuromodulin and F1), which has been implicated in different forms of synaptical plasticity in the brain such as neurite outgrowth, hippocampal LTP and neurotransmitter release, and the postsynaptic protein neurogranin (a.k.a. RC3, BICKS and p17) which function remains to be determined. This review will focus on the protein kinase C activity in pre-and postsynaptic compartment during the early phase of LTP and the possible involvement of its substrates B-50 and neurogranin.

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Characterization of a 7.5-kDa Protein Kinase C Substrate (RC3 Protein, Neurogranin) from Rat Brain

Cited by 91 publications

References 0 publications

Involvement of neurogranin in the modulation of calcium/calmodulin-dependent protein kinase II, synaptic plasticity, and spatial learning: A study with knockout mice

Involvement of neurogranin in the modulation of calcium/calmodulin-dependent protein kinase II, synaptic plasticity, and spatial learning: A study with knockout mice

Glutathiolation of Proteins by Glutathione DisulfideS-Oxide Derived from S-Nitrosoglutathione

Long-term potentiation and synaptic protein phosphorylation

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