The common structure and activities of four subspecies of rat brain protein kinase C family

Kikkawa, Ushio; Ogita, Kouji; Ono, Yoshitaka; Asaoka, Yoshinori; Shearman, Mark S.; Fujii, Tomoko; Ase, Katsuhiko; Sekiguchi, Koichi; Igarashi, Koichi; Nishizuka, Yasutomi

doi:10.1016/0014-5793(87)80291-0

Cited by 130 publications

(60 citation statements)

References 18 publications

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“…Studies conducted with both modal monothiols and dithiols indicate that cobalt and zinc compete for thiolates. The PKC zinc-thiolate motif is coordinated by a CXXC sequence in which two cysteine residues are separated by two other amino acids (25,60). This is a highly redox-active center that is present in the thioredoxin family of proteins (61).…”

Section: Discussionmentioning

confidence: 99%

A Direct Redox Regulation of Protein Kinase C Isoenzymes Mediates Oxidant-induced Neuritogenesis in PC12 Cells

Gopalakrishna

Gundimeda

Schiffman

et al. 2008

Journal of Biological Chemistry

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In this study, we have used the PC12 cell model to elucidate the mechanisms by which sublethal doses of oxidants induce neuritogenesis. The xanthine/xanthine oxidase (X/XO) system was used for the steady state generation of superoxide, and CoCl 2 was used as a representative transition metal redox catalyst. Upon treatment of purified protein kinase C (PKC) with these oxidants, there was an increase in its cofactor-independent activation. Redox-active cobalt competed with the redoxinert zinc present in the zinc-thiolates of the PKC regulatory domain and induced the oxidation of these cysteine-rich regions. Both CoCl 2 and X/XO induced neurite outgrowth in PC12 cells, as determined by an overexpression of neuronal marker genes. Furthermore, these oxidants induced a translocation of PKC from cytosol to membrane and subsequent conversion of PKC to a cofactor-independent form. Isoenzyme-specific PKC inhibitors demonstrated that PKC⑀ plays a crucial role in neuritogenesis. Moreover, oxidant-induced neurite outgrowth was increased with a conditional overexpression of PKC⑀ and decreased with its knock-out by small interfering RNA. Parallel with PKC activation, an increase in phosphorylation of the growth-associated neuronal protein GAP-43 at Ser 41 was observed. Additionally, there was a sustained activation of extracellular signal-regulated kinases 1 and 2, which was correlated with activating phosphorylation (Ser 133 ) of cAMP-responsive element-binding protein. All of these signaling events that are causally linked to neuritogenesis were blocked by antioxidant N-acetylcysteine (both L and D-forms) and by a variety of PKC-specific inhibitors. Taken together, these results strongly suggest that sublethal doses of oxidants induce neuritogenesis via a direct redox activation of PKC⑀.Understanding the signaling mechanisms involved in neuritogenesis resulting from a compensatory response to injury is crucial to the development of therapeutic agents for recovery after spinal cord and traumatic brain injuries (1, 2). The study of neuritogenesis requires the identification of molecular targets using a suitable experimental model. One of the best characterized cellular models for studying the neuronal pathways involved in neuritogenesis is the rat pheochromocytoma cell line PC12 (3). This cell line continues to be an important model system for the study of cell signaling mechanisms induced by a variety of stimuli, including neurotrophins, hormones, and oxidants (4, 5).In addition to damage from mechanical forces, spinal cord and traumatic brain injuries may result from secondary mechanisms involving ischemia, excitotoxicity, cytokines, and an infiltration of neutrophils at the site of injury (6, 7). The inflammatory response results in the generation of reactive oxygen species (ROS), 2 including superoxide and hydrogen peroxide, which can cause oxidative damage to tissues and lead to cell death (8). Paradoxically, sublethal doses of oxidants can also induce a variety of cellular processes, including cell growth, adhesion, inva...

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Section: Discussionmentioning

confidence: 99%

A Direct Redox Regulation of Protein Kinase C Isoenzymes Mediates Oxidant-induced Neuritogenesis in PC12 Cells

Gopalakrishna

Gundimeda

Schiffman

et al. 2008

Journal of Biological Chemistry

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“…PKM, but not PKC, was found exclusively in brain. The only other PKC isozymes expressed specifically in nervous tissue are the conventional PKC␥ (29) and a form of the novel PKC (30). PKM and PKC were also differentially expressed in distinct brain regions, with PKM the major form in forebrain.…”

Section: Pkm Is Expressed Exclusively In Brain-vertebrates Ex-mentioning

confidence: 99%

Protein Kinase Mζ Synthesis from a Brain mRNA Encoding an Independent Protein Kinase Cζ Catalytic Domain

Hernández

Blace

Crary

et al. 2003

Journal of Biological Chemistry

254

295

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Protein kinase M (PKM) is a newly described form of PKC that is necessary and sufficient for the maintenance of hippocampal long term potentiation (LTP) and the persistence of memory in Drosophila. PKM is the independent catalytic domain of the atypical PKC isoform and produces long term effects at synapses because it is persistently active, lacking autoinhibition from the regulatory domain of PKC. PKM has been thought of as a proteolytic fragment of PKC. Here we report that brain PKM is a new PKC isoform, synthesized from a PKM mRNA encoding a PKC catalytic domain without a regulatory domain. Multiple -specific antisera show that PKM is expressed in rat forebrain as the major form of in the near absence of full-length PKC. A PKC knockout mouse, in which the regulatory domain was disrupted and catalytic domain spared, still expresses brain PKM, indicating that this form of PKM is not a PKC proteolytic fragment. Furthermore, the distribution of brain PKM does not correlate with PKC mRNA but instead with an alternate RNA transcript thought incapable of producing protein. In vitro translation of this RNA, however, generates PKM of the same molecular weight as that in brain. Metabolic labeling of hippocampal slices shows increased de novo synthesis of PKM in LTP. Because PKM is a kinase synthesized in an autonomously active form and is necessary and sufficient for maintaining LTP, it serves as an example of a link coupling gene expression directly to synaptic plasticity. LTP1 is a persistent enhancement of synaptic transmission widely studied as a physiological model of memory (1). LTP can be divided into two phases: induction, which triggers the potentiation, and maintenance, which sustains it over time. Many molecules have been implicated in LTP induction, which is initiated by the activation of N-methyl-D-aspartate (NMDA) receptors and involves several protein kinases (2). In contrast, very little is known about the molecular mechanism of maintenance. Recently, however, a specific, autonomously active form of the atypical PKC isozyme (3, 4), PKM, has been found both necessary and sufficient for maintaining LTP (5-7). Overexpression of PKM also prolongs memory in Drosophila melanogaster, suggesting it is part of an evolutionarily conserved molecular mechanism for memory storage (8).The unique role of PKM in LTP maintenance is due, in part, to its unusual structural and enzymatic properties as an autonomously active kinase. PKM consists of the independent catalytic domain of a PKC isoform (5). PKC isoforms are divided into three classes: conventional, novel, and atypical (reviewed in Refs. 9 -11). Each isoform is a single polypeptide consisting of an N-terminal regulatory domain and a C-terminal catalytic domain linked by a hinge (Fig. 1A, left). The regulatory domain contains binding sites for second messengers and an autoinhibitory pseudosubstrate sequence, which interacts with and blocks the active site of the catalytic domain. Second messengers stimulate PKC by binding to the regulatory domain, translocating th...

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“…The amino acid sequences of protein kinases were obtained from the following sources : human G-kinase, Sandberg et al (1989); human A-kinase Cu, Madonado and Hanks (1988); human A-kinase C/3 and Cy, Beebe et al (1990); and rat protein kinase C isozymes, Kikkawa et al (1987) and Knopf et al (1986). These were aligned according to Hanks and Quinn (1991) using the LINEUP option of the Wisconsin Genetics Computer Group (GCG) suite of programs (Devereux et al, 1984).…”

Section: Sequence Alignmentsmentioning

confidence: 99%

Stimulation of cGMP‐dependent protein kinase Iα by a peptide from its own sequence

Huggins¹,

Ganzhorn²,

Saudek³

et al. 1994

European Journal of Biochemistry

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The structure of cGMP-dependent protein kinase Ia-(546 -576)-peptide amide (peptide-546) and its effects on cGMP-dependent protein kinase Ia (G-kinase) have been studied. By primary sequence analysis and analogy to a peptide that stimulates protein kinase C, peptide-546 was predicted to form part of the proteidpeptide binding site of G-kinase, and it was proposed that it would stimulate the enzyme by interaction with an autoinhibitory site. The portion of CAMP-dependent protein kinase analogous to peptide-546 forms part of the peptide substrate binding site, interacting with the peptide inhibitor residues ArgP-' and PheP-" (where p is the pseudophosphorylation site), through residues at positions corresponding to Glu4, ProlO and Serl3 in peptide-546. Peptide-546 is a reasonably potent G-kinase activator, increasing the turnover number with the peptide substrate ArgLys-Arg-Ser-Arg-Lys-Glu by about threefold with an activation constant that is about fivefold lower than the K,,, value of this peptide substrate.Peptide-546 does not appear to change the affinity of the enzyme for the above substrate, ATP or cGMP and does not affect the binding of ['HIcGMP to G-kinase. The activation does not seem to result from an interaction between peptide-546 and peptide substrates, and a kinetic scheme is proposed which is compatible with an action of peptide-546 on G-kinase independent of substrates. The activation is additive with that given by cGMP and causes the enzyme to enter a hitherto unrecognised superactive state. Peptide conformation has been monitored in mixed 2,2,2-trifluoroethanol/H,O solvents by circular dichroism: helical structure is observed in these mixtures when the 2,2,2-trifluoroethanol content is above 25%. The structure is lost only gradually on raising the temperature to 80°C with no clear melting transition.Assignment of the resonances in the 'H-NMR spectrum has allowed the identification of elements of secondary structure from detected nuclear Overhauser effects. In particular, a helical segment from Met18 to Arg26 is observed. The four proline residues (ProlO, Proll, Pro15 and Prol7) are all seen to be in the trans conformation, although additional, weaker peaks in the spectra may correspond to a minor conformer in which one or more of the prolines is in a cis conformation. The N-terminal residues are less structured but show some helical character.Calculation of the structure of the peptide using nuclear Overhauser effects (NOES) as distance constraints in distance geometry and molecular dynamics simulations revealed a well defined helical structure in the C-terminal region preceded by a less well defined proline-rich segment and Nterminal region. On the basis of these results, peptide-546 may adopt a conformation similar to that expected by analogy with CAMP-dependent protein kinase, and we propose that residues 546-576 of G-kinase are involved in enzyme autoinhibition.cGMP-dependent protein kinase is an enzyme involved in cellular signalling as a receptor protein for cGMP in Correspondence to J. P...

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The common structure and activities of four subspecies of rat brain protein kinase C family

Cited by 130 publications

References 18 publications

A Direct Redox Regulation of Protein Kinase C Isoenzymes Mediates Oxidant-induced Neuritogenesis in PC12 Cells

A Direct Redox Regulation of Protein Kinase C Isoenzymes Mediates Oxidant-induced Neuritogenesis in PC12 Cells

Protein Kinase Mζ Synthesis from a Brain mRNA Encoding an Independent Protein Kinase Cζ Catalytic Domain

Stimulation of cGMP‐dependent protein kinase Iα by a peptide from its own sequence

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