• Title, Summary, Keyword: protein kinase A

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Immunocytochemical Localization Qf raf Protein Kinase in Cerebrum of Geoclemys reevesii (Gray) (남생이(Geoclemys reevesii) 대뇌에 있어서 raf Protein Kinase의 면역세포화학적 분포)

  • 최원철;문현근
    • The Korean Journal of Zoology
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    • v.33 no.2
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    • pp.141-151
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    • 1990
  • Raf protein kinases and protein kinase C belong to serine/threonine-specific proteins in the cytoplasin, and are similar to each other in functional structure and the aspect of the distribution of celI. The distribution of raf protein kinase in the cerebrum of Geoclemys reevesfi as studied by using the antibodies against a-raf and c-raf protein kinase which induce the expression of raf fainily oncogenes. In general, raf protein kinases were distributed in such restricted regions as the general pallium, hippocampal formation, pdmordiuin hippocampi,nucleus of lateral olfactory tract, basal amygdaloid nucleus, and bed of stria terminalis. Immunological labeling of c-raf protein kinase was more widespread than that of a-raf. However, the intensity of the labeling of c-raf was lower than that of a-raf. The spherical cells of basal amygdaloid nucleus is a ring-like form, because only the cytoplasm was imunolabeled. Especially, c-raf protein kinase occurred in the cells which contained protein kinase C abundandy such as pyramidal cells and Purkinje cells. This suggests that a- and e-raf protein kinases may synegistically induce carclnoma with myc gene which is activated by protein kinase C.

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Immunocytochemical Localization of c-raf Protein Kinase in EC-4 Cell (EC-4 세포에 있어서 c-raf Protein Kinase의 면역세포화학적 위치)

  • 최원철
    • The Korean Journal of Zoology
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    • v.33 no.3
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    • pp.266-275
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    • 1990
  • c-raf protein kinase, a kind of oncogene, is a cytopiasmic serine / threonine-specific protein and is activated by mitogenic or oncogenic signals. The strncture and functions of c-raf protein kinase are considered very similar to those of protein kinase C. Using immunocytochemical approach, the time course of singal transduction of c-raf protein kinase in EC-4 cell was examined with 12-0-tetradecanoylphorbol-13-acetate (TPA) as tumor promotor and plateletderived growth factor (PDGF) as mitogenic factor. Immunoreactive c-raf was initially bound to the perinuclear membrane and then moved into the nucleus. The effect of the long-term treatment with TPA or PDGF was taken place down regulation at different time point. These results indicate that TPA and PDGF give rise to the translocation of c-raf protein kinase through the two different pathways.

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The Effects of Chronic Carbamazepine Administration on Protein Kinase A and Protein Kinase C Activities in Rat Brain (카바마제핀 장기 투여가 백서(白鼠) 뇌의 Protein Kinase A와 Protein Kinase C 활성도에 미치는 영향)

  • Rheem, Doo-Won;Kim, Leen;Suh, Kwang-Yoon
    • Korean Journal of Biological Psychiatry
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    • v.5 no.2
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    • pp.227-234
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    • 1998
  • Objective : Many evidences suggest that patients with bipolar disorder have functional abnormalities in their postreceptor signal transduction pathways, and mood stabilizing effect of lithium is exerted by modulating this dysfunctioning system. Carbamazepine, an antiepileptic agent, is also known to be effective in the treatment and prevention of bipolar disorder. But the precise mechanism of action of the drug is still poorly understood. This study was performed to elucidate the possible therapeutic mechanism of carbamazepine. Method : The effects of chronic carbamazepine administration on protein kinase A and protein kinase C activities in frontal cortex of rat brain after 2 weeks of drug administration were measured and compared with those of control subjects. Results : Mean(${\pm}SE$) value of activity(phosphate transfer ${\mu}mol/mg$ of $protein{\cdot}min$) of protein kinase A in control and test group was $0.249563{\pm}0.036$ and $0.539853{\pm}0.078$, and that of protein kinase C was $0.654817{\pm}0.053$ and $1.146205{\pm}0.052$ respectively, being increased in test group. And differences between the two groups were statistically significant for both enzymes(protein kinase A ; p<0.01, protein kinase C ; p<0.001). Conclusion : These results show that chronic carbamazepine administration increases protein kinase A and C activities, and concerning the possible mode of therapeutic action in bipolar disorder it is suggested that enhanced enzymes phosphorylate receptor-G-protein-effector complexes to dampen hyperfunctioning neuronal activity and thus stabilize the system.

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Protein Kinase C (PKC) in Cellular Signalling System: Translocation of Six Protein Kinase C Isozymes in Human Prostate Adenocarcinoma PC-3 Cell Line (세포신호계에 있어서 Protein Kinase C: 사람의 전입선 adenocarcinoma PC-3 세포내의 여섯개의 Protein kinase C 동립효소의 translocation)

  • Park, Won-Chul;Ahn, Chang-Ho
    • The Korean Journal of Zoology
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    • v.36 no.4
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    • pp.439-451
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    • 1993
  • Protein kinase C isozymes in a human prostate adenocarcinoma PC-3 cell line were characterized. Immunoreactive bands and immunocytochemical stains were obsenred in PC-3 cells with antibodies raised against protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$, and ζ types, respectively. Protein kinase C ${\alpha}$ corresponded to a immunoreactive band at a molecular weight of 80,000-dalton, whereas molecular weights of other immunoreactive isozvmes of protein kinase C were detected at 68,000-dalton. Protein kinHse C $\delta$ and ζ antibodies detected additional bands at 55,000-dalton and 80,000-dalton, respectively Immunocvtochemical study confirmed the results of the immunoblotting experiments qualitatively: all six protein kinase C isozymes were detected in the cytoplasm of PC-3 cells. Translocation of protein kinase C in PC-3 cells were also examined with phorbol 12-myristate 13-acetate (PMA), bryostatin 2, diolein, and 1-oleoyl-2-acetyl glycerol (OAG). Differential reactions of protein kinase C isozvmes to these activators were obsenred. When PC-3 cells were treated with 10mM bryostatin 2, protein kinase C isozyme u was translocated into the nucleus, whereas s type was translocated into the plasma membrane and the nucleus. Protein kinase C ${\alpha}$ and ζ types were translocated into the nucleus following the treatment with 101M diolein, whereas protein kinase C ${\alpha}$, ${\beta}$, ${\gamma}$, and $\varepsilon$ types were translocated into the nucleus by the treatment with 10mM OAG. Protein kinase C ${\alpha}$ and $\varepsilon$ types were translocated into the nucleus in the presence of 100nM PMA. Protein kinase C $\delta$ type was translocated to the nuclear membrane by these activators, however, only PMA-induced translocation was inhibited by protein kinase C inhibitor, 1-(5-isoquinolinesulfonyll-2-methvlpiperazine dihvdrochloride (H7) . H7 inhibited translocation of protein kinase C ${\alpha}$ type induced by PMA, ${\beta}$ type by OAG and s type by PMA and OAG, whereas it did not affect translocations induced by bryostatin and diolein, respectively. These results suggest that there exist six isoformes of protein kinase C (${\alpha}$, ${\beta}$, ${\gamma}$, $\delta$, $\varepsilon$ and ζ types) in PC-3 cells and that each of these isozvmes distinctivelv reacts to bryostatin, diolein, OAG and PMA, in part due to an altered molecular size and conceivably discrete binding site(s).

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Action of Protein Kinase A and C Activators on Germinal Vesicle Breakdown and One-Cell Embryos in the Mouse (생쥐 GV난자와 1-세포기 배아의 핵막붕괴에 미치는 Protein Kinase A와 C의 작용)

  • 이대기;김경진;조완규
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.153-162
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    • 1989
  • Expedments were perfonned to examine the role of cAMP-dependent protein kinase (PK-A) and diacylglycerol-dependent protein kinase (PK-C) during the meiodc resumption and the first mitotic cell cycle of mouse embryogenesis. Mejoric GV oocytes and one-cell embryos derived from in vitro fertilization were cultured in vitro, and morphological changes in response to activators of PK-A and PK-C were examined. Treatments with a membrane-permeable cAMP analog, dbcAMP (0.1 mg/mi), phosphodiesterase inhibitor, IBMX (0.1 mM), biologically active phorbol ester, WA (10 nglmi), or a synthetic diacylglycerol, sn-diC8 inhibited resumption of melosis. Combination of PK-A and PK-C activator brought about furiher inhibition. On the contrary, dbcAMP (0.1 mg/mi), IBMX (0.2 mM), WA (10 nglml), and sn-diC8 (0.5 mM) did not inhibit pronucleus membrane breakdown (PNBD) when added S or G2 phase of cell cycle. However, activators of PK-C inhibited cleavage of one-cefl embryos. This result indicates that the action mechanism of PK-A and PK-C on dissolution of nuclear membrane in primary meiotic arrest oocytes may be different from that of mitotic one-cell embryos.

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The Identification of Type II DNA Topoisomerase-Associated Protein Kinase Activity from Regenerating Rat Liver (재생 쥐간에서 분리한 DNA topoisomerase II에 결합된 protein kinase 활성)

  • 이치건;박세호;남궁록;김찬길;박상대
    • The Korean Journal of Zoology
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    • v.36 no.3
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    • pp.367-372
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    • 1993
  • We have found a protein kinase activity that is tightly associated with type II DNA topoisomerase purified from regenerating rat liver. The activities of protein kinase and topoisomerase II were not separable when the enzyme was subjected to analytical chromatographies (Hydroxyapatite, phosphocellulose, and double strand DNA cellulose) and glycerol gradient sedimentation. The kinase activity from purified rat topoisomerase II was also inactivated by the topoisomerase II inhibitors such as N-ethylmaleimide or novobiocin. The evidences, however, do not rule out a possibility that the kinase activity resides in a polypeptide other than the topoisomerase II protein. The topoisomerase II-associated protein kinase required Mg++ for its activity, and this requirement was not substituted by any other mono- or divalent ions. Histone H1 act as a strong stimulator and a good substrate for the kinase activity and other histones and ${\alpha}$-casein could not substitute the effect of histone H1.

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Molecular Characterization of a Protein Kinase Gene in Chiness Cabbage(Brassica campestrics subsp. napus var. pekinensis)

  • Jeong, Sang-Ho;Ahn, Ji-Hoon;Lee, June-Seung;Lee, Jong-Seob
    • Animal cells and systems
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    • v.1 no.1
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    • pp.135-142
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    • 1997
  • Random sequencing of expressed sequence tags in roots of Chinese cabbage led to isolation of a partial cDNA clone, BR77, which encoded a putative protein kinase. Using the BR77 cDNA as a probe, we isolated a full-length cDNA encoding the Brassica campestris protein kinase 1 (Bcpk1). The Bcpt1 cDNA contained one open reading frame encoding a polypeptide of 439 amino acids. The putative polypeptide consisted of a short N-terminal region and a protein kinase catalytic domain. The catalytic domain of Bcpkl showed a high homology to cAMP- and calcium- phospholipid-dependent subfamilies of serine/threonine protein kineses. Eleven major catalytic domains in protein kineses were well conserved in Bcpk1. However, Bcpk1 contained a unique nonhomologous intervening sequence between subdomains VII and VIII, which was not found in protein kineses of animals and lower eukaryotes. Genomic DNA gel blot analysis showed that Bcpt1 genes might be present as three copies in the Chinese cabbage genome. These imply that Bcpk1 belongs to a plant-specific serine/threonine protein kinase subfamily.

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Three Protein Kinases from the Etiolated Oat Seedlings Phosphorylate Oat Phytochrome A In Vitro

  • Park, Young-Il;Kim, Jae-Hun;Lee, Jae-Deok;Kim, Yong-Woo;Kim, In-Soo
    • BMB Reports
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    • v.31 no.3
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    • pp.221-226
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    • 1998
  • Phosphorylation of phytochrome may play important functional roles to control plant photomorphogenesis. Many attempts have failed to identify the protein kinase that phosphorylates phytochrome in vivo. It has been reported that a polycation-stimulated protein kinase activity was associated with the purified phytochrome. However, it is not known if the kinase activity is an intrinsic property of phytochrome or whether it comes from a contaminant of the purified phytochrome. In the present study, three protein kinases that phosphorylate phytochrome have been identified from etiolated oat seedlings. A polycationstimulated protein kinase that had very similar enzymatic properties with that associated with the purified phytochrome was identified in the cytosolic extract. It phosphorylated several contaminant proteins in the kinase preparation as well as phytochrome and had a broad substrate specificity. A CK II-type protein kinase phosphorylated phytochrome and the exogenously added casein. It is likely that this kinase may not be a feasible candidate for the kinase phosphorylating phytochrome in vivo since the content of the kinase seemed to well exceed the content of phytochrome in the etiolated oat seedlings. Another protein kinase that had unique enzymatic properties phosphorylated phytochrome very specifically and seemed to be present in a small quantity in the etiohlted seedlings. It is expected that one of three kinases may be responsible for the phytochrome phosphorylation in vivo.

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New Alternative Splicing Isoform and Identification of the Kinase Activity of N-Terminal Kinase-Like Protein (NTKL)

  • Merlin, Jayalal L.P.
    • Journal of the Chosun Natural Science
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    • v.6 no.4
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    • pp.234-243
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    • 2013
  • N-terminal kinase-like (NTKL) protein was initially identified as a protein binding to protein kinase B (PKB, also known as Akt). Though NTKL-BP1 (NTKL-binding protein 1) has been identified as an NTKL binding protein, its functions related to binding have not yet been elucidated. Here, a new alternative spliced variant of NTKL and its association with integrin ${\beta}1$ is described, in addition to the kinase activity of NTKL and its substrate candidates. Although the phosphorylation of the candidates must be further confirmed using other experimental methods, the observation that NTKL can phosphorylate ROCK1, DYRK3, and MST1 indicates that NTKL may act as a signaling protein to regulate actin assembly, cell migration, cell growth, and to facilitate differentiation and development in an integrin-associated manner.

Changes in Kinetic Properties of $Ca^{2+}$/Calmodulin-Dependent Protein Kinase la Activated by $Ca^{2+}$/Calmodulin-Dependent Protein Kinase I Kinase (칼슘/칼모듈린-의존성 단백질 키나아제 I 키나아제에 의한 칼슘/칼모듈린-의존성 단백질 키나아제 Ia의 활성화에 따른 효소반응 특성의 변화)

  • Cho, Jung-Sook
    • YAKHAK HOEJI
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    • v.41 no.6
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    • pp.773-781
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    • 1997
  • The activity of $Ca^{2+}$calmodulin (CaM)-dependent protein kinase Ia (CaM kinase Ia) is shown to be regulated through direct phosphorylation by CaM kinase I kinase (CaMK IK). In the present study, three distinct CaMKIK peaks were separated from Q-Sepharose colunm chromatography of pig brain homogenate using a Waters 650 Protein Purification System. The purified CaMKIK from the major peak potently and rapidly enhanced CaM kinase Ia activity, reaching a maximal stimulation within 2min at the concentrations of 12-15nM. The activated state of CaM kinase Ia is characterized by a markedly enhanced $V_{max}4 as well as significantly decreased $K_m\;and\;K_a$ values toward peptide substrate and CaM, respectively. These observations suggest the activation process of CaM kinase Ia. The phosphorylation of CaM kinase Ia by CaMKIK may induce its conformational change responsible for the alterations in the kinetic properties, which ultimately leads to the rapid enzyme activation.

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