Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.11.13 (protein kinase C)
 49,245 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The effects on neuronal nitric oxide synthase (NOS) of protein kinase C (PKC) activation in rat cerebellar slices and of in vitro phosphorylation by PKC were compared. Incubation of slices with 1-aminocyclopentane-1,3-trans-dicarboxylic acid (trans-ACPD) or phorbol myristate acetate (PMA) in the presence of okadaic acid (OA) shifted the calcium sensitivity of neuronal NOS in the homogenate or in the cytosolic fraction. trans-ACPD promoted translocation of PKC activity to the particulate fraction in the slices. PMA in the presence of OA enhanced phosphorylation of GAP43 protein in the slices. These results ensured that both treatments activated PKC in the slice. However, when neuronal NOS in the slice treated with PMA and OA, in which GAP43 phosphorylation was detected, was immunoprecipitated by a specific antibody, no indication of neuronal NOS phosphorylation was obtained. Nevertheless, PKC phosphorylated partially purified neuronal NOS in vitro. Phosphorylated neuronal NOS showed greater activity than unphosphorylated NOS, but their calcium sensitivity was identical. These data indicated that neuronal NOS is not susceptible to PKC-dependent phosphorylation in cerebellar slices and that the calcium-sensitivity shift of neuronal NOS takes place without direct phosphorylation of neuronal NOS, suggesting the involvement of unknown proteins whose phosphorylation would regulate the calcium sensitivity of neuronal NOS in the cerebellum.
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PMID:Differential effects of protein kinase C on neuronal nitric oxide synthase activity in rat cerebellar slices and in vitro. 881 26

Recently, we reported that in vitro and in vivo persistent infection of neurons by lymphocytic choriomeningitis virus (LCMV) downregulated GAP43 expression, a protein involved in neuronal plasticity associated with learning and memory. Here, we investigated the transcriptional and posttranscriptional events involved. Persistent LCMV infection of PC12 cells (PC12Pi) caused reduced levels of GAP43 steady-state mRNA when compared to uninfected PC12 cells. In addition, an increase in the steady-state levels of GAP43 mRNA observed in PC12 cells in response to nerve growth factor (NGF) was abrogated in PC12Pi cells. Nuclear run-on analysis revealed that the rate of GAP43 transcription was reduced threefold in PC12Pi cells compared to uninfected PC12 cells. Moreover, analysis of the half-life of GAP43 mRNA indicated that NGF-mediated stabilization of GAP43 transcripts was significantly diminished in PC12Pi cells. Treatment of PC12Pi cells with basic fibroblast growth factor, dibutyryl cyclic AMP, and 12-o-tetradecanoyl-phorbol-13-acetate, a potent activator of protein kinase C, did not increase the GAP43 mRNA steady-state level, suggesting that LCMV infection interferes with a step downstream from protein kinases A and C in the NGF signal transduction pathway.
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PMID:Viral persistent infection affects both transcriptional and posttranscriptional regulation of neuron-specific molecule GAP43. 914 70

The dynamic properties of the cell cortex and its actin cytoskeleton determine important aspects of cell behavior and are a major target of cell regulation. GAP43, myristoylated alanine-rich C kinase substrate (MARCKS), and CAP23 (GMC) are locally abundant, plasmalemma-associated PKC substrates that affect actin cytoskeleton. Their expression correlates with morphogenic processes and cell motility, but their role in cortex regulation has been difficult to define mechanistically. We now show that the three proteins accumulate at rafts, where they codistribute with PI(4,5)P(2), and promote its retention and clustering. Binding and modulation of PI(4, 5)P(2) depended on the basic effector domain (ED) of these proteins, and constructs lacking the ED functioned as dominant inhibitors of plasmalemmal PI(4,5)P(2) modulation. In the neuron-like cell line, PC12, NGF- and substrate-induced peripheral actin structures, and neurite outgrowth were greatly augmented by any of the three proteins, and suppressed by DeltaED mutants. Agents that globally mask PI(4,5)P(2) mimicked the effects of GMC on peripheral actin recruitment and cell spreading, but interfered with polarization and process formation. Dominant negative GAP43(DeltaED) also interfered with peripheral nerve regeneration, stimulus-induced nerve sprouting and control of anatomical plasticity at the neuromuscular junction of transgenic mice. These results suggest that GMC are functionally and mechanistically related PI(4,5)P(2) modulating proteins, upstream of actin and cell cortex dynamics regulation.
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PMID:GAP43, MARCKS, and CAP23 modulate PI(4,5)P(2) at plasmalemmal rafts, and regulate cell cortex actin dynamics through a common mechanism. 1087 Dec 85

The ability to respond rapidly to changes in tonicity is crucial for cellular and organismal survival. Sensors of osmotic stress are beginning to be discovered. For example, results from expression cloning in a heterologous system have implicated GAP43 as a component of a peripheral nervous system sensor of hypotonicity. These results and the role of lipid rafts, protein kinase C, and members of the phospholipase C-delta family are discussed in the context of cellular responses to osmotic stress. Calcium is also involved in the osmotic stress response, and both intracellular calcium released through inositol trisphosphate receptors and extracellular calcium transported through TRPV4 (a member of the transient receptor potential family) may contribute.
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PMID:Of rafts and moving water. 1296 85

The possibility of coexistence of different subtypes of membrane lipid rafts has been investigated in cerebellar granule cells, by submitting detergent-resistant membrane fractions to immunoprecipitation. Among the proteins and lipids present in detergent-resistant fractions, almost all Prion protein, GAP43 and PKC were present in the immunoprecipitate obtained with anti-GAP43 or anti-Prion protein antibody at 4 degrees C, together with a small fraction of cholesterol and sphingolipids, suggesting that they belong to a distinct subset of membranes. On the contrary, all Fyn and almost all MARCKS remained in the supernatant. Fluorescence microscopy experiments showed that Fyn and Prion protein were mostly not colocalized within a single neuron. Our results suggest that granule cells membranes contains different subtypes of detergent-resistant fractions, possibly deriving from different lipid rafts.
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PMID:Immunoseparation of Prion protein-enriched domains from other detergent-resistant membrane fractions, isolated from neuronal cells. 1474 57

Patterned visual activity, acting via NMDA receptors, refines developing retinotectal maps by shaping individual retinal arbors. Because NMDA receptors are postsynaptic but the retinal arbors are presynaptic, there must be retrograde signals generated downstream of Ca(++) entry through NMDA receptors that direct the presynaptic retinal terminals to stabilize and grow or to withdraw. This review defines criteria for retrograde synaptic messengers, and then applies them to the leading candidates: nitric oxide (NO), brain-derived neurotrophic factor (BDNF), and arachidonic acid (AA). NO is not likely to be a general mechanism, as it operates only in selected projections of warm blooded vertebrates to speed up synaptic refinement, but is not essential. BDNF is a neurotrophin with strong growth promoting properties and complex interactions with activity both in its release and receptor signaling, but may modulate rather than mediate the retrograde signaling. AA promotes growth and stabilization of synaptic terminals by tapping into a pre-existing axonal growth-promoting pathway that is utilized by L1, NCAM, N-cadherin, and FGF and acts via PKC, GAP43, and F-actin stabilization, and it shares some overlap with BDNF pathways. The actions of both are consistent with recent demonstrations that activity-driven stabilization includes directed growth of new synaptic contacts. Certain nondiffusible factors (synapse-specific CAMs, ephrins, neurexin/neuroligin, and matrix molecules) may also play a role in activity-driven synapse stabilization. Interactions between these pathways are discussed.
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PMID:Activity-driven sharpening of the retinotectal projection: the search for retrograde synaptic signaling pathways. 1500 31

The interactions of cells with their environment involve regulated actin-based motility at defined positions along the cell surface. Sphingolipid- and cholesterol-dependent microdomains (rafts) order proteins at biological membranes, and have been implicated in most signalling processes at the cell surface. Many membrane-bound components that regulate actin cytoskeleton dynamics and cell-surface motility associate with PtdIns(4,5)P(2)-rich lipid rafts. Although raft integrity is not required for substrate-directed cell spreading, or to initiate signalling for motility, it is a prerequisite for sustained and organized motility. Plasmalemmal rafts redistribute rapidly in response to signals, triggering motility. This process involves the removal of rafts from sites that are not interacting with the substrate, apparently through endocytosis, and a local accumulation at sites of integrin-mediated substrate interactions. PtdIns(4,5)P(2)-rich lipid rafts can assemble into patches in a process depending on PtdIns(4,5)P(2), Cdc42 (cell-division control 42), N-WASP (neural Wiskott-Aldrich syndrome protein) and actin cytoskeleton dynamics. The raft patches are sites of signal-induced actin assembly, and their accumulation locally promotes sustained motility. The patches capture microtubules, which promote patch clustering through PKA (protein kinase A), to steer motility. Raft accumulation at the cell surface, and its coupling to motility are influenced greatly by the expression of intrinsic raft-associated components that associate with the cytosolic leaflet of lipid rafts. Among them, GAP43 (growth-associated protein 43)-like proteins interact with PtdIns(4,5)P(2) in a Ca(2+)/calmodulin and PKC (protein kinase C)-regulated manner, and function as intrinsic determinants of motility and anatomical plasticity. Plasmalemmal PtdIns(4,5)P(2)-rich raft assemblies thus provide powerful organizational principles for tight spatial and temporal control of signalling in motility.
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PMID:Spatial control of actin-based motility through plasmalemmal PtdIns(4,5)P2-rich raft assemblies. 1564 36

The inner leaflet of a typical mammalian plasma membrane contains 20-30% univalent PS (phosphatidylserine) and 1% multivalent PtdIns(4,5)P(2). Numerous proteins have clusters of basic (or basic/hydrophobic) residues that bind to these acidic lipids. The intracellular effector CaM (calmodulin) can reverse this binding on a wide variety of proteins, including MARCKS (myristoylated alanine-rich C kinase substrate), GAP43 (growth-associated protein 43, also known as neuromodulin), gravin, GRK5 (G-protein-coupled receptor kinase 5), the NMDA (N-methyl-D-aspartate) receptor and the ErbB family. We used the first principles of physics, incorporating atomic models and the Poisson-Boltzmann equation, to describe how the basic effector domain of MARCKS binds electrostatically to acidic lipids on the plasma membrane. The theoretical calculations show the basic cluster produces a local positive electrostatic potential that should laterally sequester PtdIns(4,5)P(2), even when univalent acidic lipids are present at a physiologically relevant 100-fold excess; four independent experimental measurements confirm this prediction. Ca(2+)/CaM binds with high affinity (K(d) approximately 10nM) to this domain and releases the PtdIns(4,5)P(2). MARCKS, a major PKC (protein kinase C) substrate, is present at concentrations comparable with those of PtdIns(4,5)P(2) (approx. 10 microM) in many cell types. Thus MARCKS can act as a reversible PtdIns(4,5)P(2) buffer, binding PtdIns(4,5)P(2) in a quiescent cell, and releasing it locally when the intracellular Ca(2+) concentration increases. This reversible sequestration is important because PtdIns(4,5)P(2) plays many roles in cell biology. Less is known about the role of CaM-mediated reversible membrane binding of basic/hydrophobic clusters for the other proteins.
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PMID:Reversible - through calmodulin - electrostatic interactions between basic residues on proteins and acidic lipids in the plasma membrane. 1564 42

Changes in the composition of cell fractions, and in particular of detergent-resistant membranes (DRM) isolated from cultured rat cerebellar granule cells, were taken as possible changes in lipid raft composition during a signal transduction event. After activation of protein kinase C (PKC) with phorbol esters (PMA) or glutamate, the content of PKC and of proteins highly enriched (GAP43, Fyn, and PrP(c)) or not (MARCKS) in DRM was followed. PKC activation strongly increased its association with membranes (from 2% to 75%), causing its enrichment within DRM; the substrate GAP43, enriched in DRM, remained membrane associated, but its proportion in DRM dramatically decreased (from about 40% to 2.5%), suggesting its shift from raft to nonraft membranes, possibly as a consequence of phosphorylation by PKC. The distribution of Fyn and PrP(c) (DRM-enriched) and of MARCKS (present mainly outside DRM) did not change. PKC activation was followed by an increase of GAP43 and MARCKS phosphorylation (about 7- and 8-fold, respectively). Noteworthy was that, after cell treatment with the lipid raft-disrupting drug methyl-beta-cyclodextrin, PKC activation occurred normally, followed by MARCKS phosphorylation, but GAP43 phosphorylation did not occur. Taken altogether, these data suggest that the integrity of lipid rafts is necessary for PKC to affect GAP43 and catalyze its phosphorylation.
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PMID:Changes in the composition of detergent-resistant membrane domains of cultured neurons following protein kinase C activation. 1708 51

In the developing visual system, correlated presynaptic activity between neighboring retinal ganglion cells (RGC) stabilizes retinotopic synapses via a postsynaptic NMDAR (N-methyl-D-aspartate receptor)-dependent mechanism. Blocking NMDARs makes individual axonal arbors larger, which underlies an unsharpened map, and also increases branch turnover, as if a stabilizing factor from the postsynaptic partner is no longer released. Arachidonic acid (AA), a candidate retrograde stabilizing factor, is released by cytoplasmic phospholipase A2 (cPLA2) after Ca(2+) entry through activated NMDARs, and can activate presynaptic protein kinase C to phosphorylate various substrates such as GAP43 to regulate cytoskeletal dynamics. To test the role of cPLA2 in the retinotectal system of developing zebrafish, we first used PED6, a fluorescent reporter of cPLA2 activity, to show that 1-3 min of strobe flashes activated tectal cPLA2 by an NMDAR-dependent mechanism. Second, we imaged the dynamic growth of retinal arbors during both local inhibition of tectal cPLA2 by a pharmacological inhibitor, arachidonic tri-fluoromethylketone, and its suppression by antisense oligonucleotides (both injected intraventricularly). Both methods produced larger arbors and faster branch dynamics as occurs with blocking NMDARs. In contrast, intraocular suppression of retinal cPLA2 with large doses of antisense oligos produced none of the effects of tectal cPLA2 inhibition. Finally, if AA is the retrograde messenger, the application of exogenous AA to the tectum should reverse the increased branch turnover caused by blocking either NMDARs or cPLA2. In both cases, intraventricular injection of AA stabilized the overall branch dynamics, bringing rates down below the normal values. The results suggest that AA generated postsynaptically by cPLA2 downstream of Ca(2+) entry through NMDARs acts as a retrograde signal to regulate the dynamic growth of retinal arbors.
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PMID:Arachidonic acid as a retrograde signal controlling growth and dynamics of retinotectal arbors. 1791 41


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