• Title/Summary/Keyword: PKC

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Effects of Bulbus Allii Macrostemi Extract on PKC activity in Pulmonary Vascular Endothelial Cells Damaged by XO/HX (산소자유기에 의해 손상된 혈관내피세포의 PKC 활성도에 미치는 해백 추출물의 효과(I))

  • Kwon Kang Beom;Lee Ho Seung;Kang Gil Seong;Kim In Seob;Ryu Do Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.2
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    • pp.443-446
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    • 2003
  • To investigate the protective effect of Bulbus Allii Macrostemi (BAM) on the damage by pulmonary vascular endothelial cells by xanthine oxidase (XO)/hypoxanthine (HX)-induced oxygen free radical, Neutral Red (NR) and protein kinase c (PKC) activity assay were used. The results were obtained as follows ; The viability of vascular endothelial cells treated with XO/HX was decreased. And activation of PKC represented a maximal increase in group treated with XO/HX for 15 mins in vasvular pulmonary endothelial cells. But pretreated groups with BAM extracts were not inhibited the increase of PKC activation by XO/HX in a dose-dependent fashion. These results show that XO/HX elicits toxic effects in cultured pulmonary vascular endothelial cells, and suggest that BAM extract is very effective in the prevention of XO/HX-induced PKC activation.

Protein kinase C와 이와 관련된 단백질 연구

  • 이재란;김진한;최명언
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.135-135
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    • 1993
  • 신호전달과정에 중요한 역할을 하고 있는 다기능 serinei/threonine 단백질인산화효소인 protein kinase C(PKC)의 연구를 위해 이 효소의 정제를 뇌에서 착수하였다 PKC의 활성측정을 myelin basic protein을 기질로 하여 20 mM Tris 완충용액 PH 7.5, 0.15 mM [${\gamma}$-$^{32}$P]ATP(3 $\times$ $10^{5}$ cpm), 0.1 mM $Ca^{2+}$, 10$\mu\textrm{g}$ phosphatidylserine과 2$\mu\textrm{g}$ diolein을 넣어 반응시켰다. 반응은 TCA로 정지시킨 후 방사성 단백질을 Millipore filter paper로 걸러 섬광 계수기로 읽었다. Cytosol PKC의 정제과정은 첫 단계에서 DEAE-cellulose를 사용하였으며, phenyl sepharose CL-4B와 protamine agarose를 연속적으로 이용하여 800배의 정제에 성공했다. SDS-PACE 상에서 80 kD로 나타났으며 순도는 95 % 이상이였다. 이를 이용 PKC의 각종 기질 연구에 착수하기 시작했으며, 이중 MBP의 인산화연구를 통한 myelin의 안정성과 MBP와의 구조 관계가 일부 수행되고 있다 연차적으로 PKC와 이와 관련된 단백질의 특성을 살피기 위해 뇌의 PKC 기질 중 cold stress를 통해 환경에 민감한 것을 찾고 있으며, 현재 autoradiography를 이용해 80 kD, 54 kD, 49 kD와 35 kD의 단백질이 연구대상이 되고있다. 그 중 49 kD는 B-50(또는 GAP43, neuromodulin이라고도 함)일 가능성이 높아 이 단백질 조절과 PKC 활성화 사이의 관계 정립이 흥미로운 과제로 대두되고 있다.다.

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Ligand-Based CoMFA Study on Pyridylpyrazolopyridine Derivatives as PKCθ Kinase Inhibitors

  • Balasubramanian, Pavithra K.;Balupuri, Anand;Cho, Seung Joo
    • Journal of Integrative Natural Science
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    • v.7 no.4
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    • pp.253-259
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    • 2014
  • Protein kinase C theta (PKC-${\theta}$) is a serine/threonine specific protein kinase. It is largely expressed in the T-cells and CD28 signaling. PKC-${\theta}$ phosphorylates diverse proteins that are involved in the various cellular signaling pathways. Activated PKC-${\theta}$ in turn activates other transcription factors that control the proliferation and differentiation of T- cells. PKC-${\theta}$ is considered to be an interesting therapeutic target due to its crucial role in the proliferation, differentiation and survival of T-cells. In the present study, we have performed ligand-based CoMFA study on a series of pyridylpyrazolopyridine derivatives as PKC-${\theta}$ inhibitors. An acceptable CoMFA model ($q^2$=0.544; ONC=4; $r^2$=0.876) was developed and validated by Bootsrapping and progressive sampling. The CoMFA contour map suggested the regions to increase the activity. Bulky substitutions in R2 position of the piperizine ring could increase the activity. Similarly positive, small substitution in the R1 position of the Pyridine ring could considerably increase the activity. Our work could assist in designing more potent PKC-${\theta}$ inhibitors of pyridylpyrazolopyridine derivatives.

Protein Kinase C-delta Stimulates Haptoglobin Secretion

  • Oh, Mi-Kyung;Park, Seon-Joo;Kim, Nam-Hoon;Kim, In-Sook
    • BMB Reports
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    • v.40 no.1
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    • pp.130-134
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    • 2007
  • Haptoglobin (Hp) is a glycoprotein that is produced by hepatic cells and secreted into the circulation. While studying the physiologic functions of Hp, we found that Hp synthesized in THP-1 monocytic cells was largely retained within cells, although Hp is considered a secretory protein. To investigate the molecular mechanism on Hp secretion in THP-1 cells, in the present study, we examined the effect of protein kinase C (PKC) on Hp secretion. When several inhibitors of PKC isoforms were tested, only Rottlerin, a specific inhibitor of PKC-$\delta$, completely blocked Hp secretion from cells to culture medium. To confirm the role of PKC-$\delta$ in Hp secretion, Hp-overexpressing COS7 cells were transiently transfected with a wild-type or a dominantnegative mutant of the PKC-$\delta$ gene. Mutant PKC-$\delta$ significantly inhibited Hp secretion, whereas the wild-type gene slightly increased Hp secretion. These results demonstrate that the PKC-$\delta$ signal is involved in Hp secretion.

Preimage Attacks on Reduced Steps of ARIRANG, HAS-160, and PKC98-Hash (ARIRANG, HAS-160, PKC98-Hash의 축소된 단계들에 대한 역상공격)

  • Hong, Deuk-Jo;Koo, Bon-Wook;Kim, Woo-Hwan;Kwon, Dae-Sung
    • Journal of the Korea Institute of Information Security & Cryptology
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    • v.20 no.2
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    • pp.3-14
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    • 2010
  • In this paper, we present the preimage attacks on step-reduced ARIRANG, HAS-160, and PKC98-Hash. We applied Aoki and Sasaki's chunk serach method which they have used in the attack on SHA-0 and SHA-1. Our attacks find the preimages of 35-step ARIRANG, 65-step HAS-160, and 80-step PKC98-Hash. Our results are the best preimage attacks for ARIRANG and HAS-160, and the first preimage attack for PKC98-Hash faster than exhaustive search.

Effect of Feeding Rubber Seed Kernel and Palm Kernel Cake in Combination on Nutrient Utilization, Rumen Fermentation Characteristics, and Microbial Populations in Goats Fed on Briachiaria humidicola Hay-based Diets

  • Chanjula, P.;Siriwathananukul, Y.;Lawpetchara, A.
    • Asian-Australasian Journal of Animal Sciences
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    • v.24 no.1
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    • pp.73-81
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    • 2011
  • Six male crossbred (Thai Native${\times}$Anglo Nubian) goats, with an average initial weight of $22{\pm}2\;kg$, were randomly assigned according to a $3{\times}2$ factorial arrangement in a $6{\times}6$ Latin square design with a 21-d period to evaluate the effect of feeding rubber seed kernel (RSK) and palm kernel cake (PKC) in combination on nutrient utilization, rumen fermentation characteristics, and nitrogen utilization. The dietary treatments were as follows: i) concentrate containing 0% RSK and 20% PKC ($T_1$), ii) 0% RSK and 30% PKC ($T_2$), iii) 20% RSK and 20% PKC ($T_3$), iv) 20% RSK and 30% PKC ($T_4$), v) 30% RSK and 20% PKC ($T_5$), and vi) 30% RSK and 30% PKC ($T_6$). During the experiment, signal hay was given on an ad libitum basis as the roughage. It was found that RSK levels and PKC levels had no interaction effects on feed intake, apparent digestibility, $NH_3$-N, blood metabolites, VFA concentrations, and nitrogen utilization, but there were interactions between RSK levels and PKC levels with respect to total DMI (kg/d) and total VFA concentrations, and goats receiving 30% RSK had lower values (p<0.05) than those receiving 0 and 20% RSK, respectively. Feeding different PKC levels did not affect (p>0.05) feed intake, digestibility, rumen fermentation patterns, blood metabolites, and nitrogen utilization. However, increasing RSK levels (>20%) resulted in a slightly lower daily DMI (% BW and g/kg $BW^{0.75}$), apparent digestibility (NDF and ADF), total N intake, and N excretion than in goats fed on 0 and 20% RSK. BUN, blood glucose, and propionate were variable among treatment and were highest in 0% RSK with the 20% PKC fed group having values which were higher than those in other groups. However, there were no differences (p>0.05) among treatments with respect to N retention, PD output, and microbial N supply. Based on this study, RSK levels up to 20% and PKC at 20-30% in concentrate could be efficiently utilized for goats fed on signal hay.

Physicochemical Properties and Antioxidant Activities of High-Sugar Fermented Pinus koraiensis Cone Extract (잣송이 추출물의 이화학적 특성 및 항산화 활성)

  • Jang, Ki-Hyo;Surh, Jeonghee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.46 no.1
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    • pp.140-145
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    • 2017
  • Pinus koraiensis cone (PKC) extract was prepared by mixing PKC with sugar at a weight ratio of 1:1 and then removing PKC after incubating the mixture at room temperature for 100 days. The resulting PKC extract was examined for its physicochemical properties and antioxidant activities using Prunus mume fruit extract, which is similar to PKC extract in terms of preparation procedure and product usage, as a control. PKC extract consisted mainly of moisture (21.76%) and sugar (70.03%), leading to high viscosity. PKC extract was apparently yellowish, and therefore its browning index and color intensity were appreciably lower than the control (P=0.0000), indicating that Maillard reaction was not facilitated during fermentation. Compared with the control, PKC extract was not appreciably different in terms of total flavonoid content, whereas its organic acid content and total reducing capacity were significantly lower. Nevertheless, its metal-chelating activity and DPPH radical scavenging activity were comparable to those of the control. In addition, SOD-like activity of PKC extract was 2-fold higher than that of 6.6 mM quercetin, which had a higher total flavonoid content than PKC extract. This indicates that PKC extract contains certain flavonoids with higher antioxidant activities than quercetin. The results show that PKC extract provides antioxidant activity as well as characteristically different sensory properties due to its higher sugar and lower organic acid contents compared with Prunus mume fruit extract.

Effects of Chitosan on the Toxicity of Environmental Pollutants (해양바이오물질이 PCB의 독성작용에 미치는 영향)

  • Lee, Hyon-Gyo;Kim, Hae-Young;Yang, Jae-Ho
    • Journal of Marine Bioscience and Biotechnology
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    • v.2 no.2
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    • pp.102-107
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    • 2007
  • Environmental contamination becomes a great public concern as our society gets industrialized rapidly. The present study examine the role of chitosan in a effort to intervene the environmental pollutant-induced toxicity. PCB-induced neurotoxicity with respect to the PKC signaling was examined. Since the developing neuron is particularly sensitive to PCB-induced neurotoxicity, we isolated cerebellar granule cells derived from 7-day old SD rats and grew cells in culture for additional 7 days to mimic PND-14 conditions. PCB showed the alteration of PKC signaling pathway. The alteration was structure-dependent. Mono-ortho-substituted congeners at a high dose showed a significant increase of total PKC activity at [$^3H$]PDBu binding assay, indicating that mono-ortho-substituted congeners are more neuroactive than non-ortho-substituted congeners in neuronal cells. PKC isoforms were immunoblotted with respective monoclonal antibodies. PKC-beta II and -epsilon were activated with mono-ortho-substituted congeners exposure. The result suggests that the position with ortho has a higher potential of altering the signaling pathway. Alteration of PKC was blocked with treatment of high molecular weight of chitosan. The study demonstrated that the ortho position in PCBs are important in assessing the structure-activity relationship. The results suggest a potential use of marine bioactive materials as a means of nutritional intervention to prevent the harmful effects of pollutant-derived toxicity.

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THE EFFECT OF PKC PATHWAY & MAPK PATHWAY ON RUNX2 TRANSCRIPTIONAL ACTIVITY (Protein kinase C 및 MAPK pathway가 Runx2의 전사 활성에 미치는 영향)

  • Kim, Eun-Jung;Kim, Hyun-Jung;Ryoo, Hyun-Mo;Kim, Hyun-Jung;Kim, Young-Jin;Nam, Soon-Hyeun
    • Journal of the korean academy of Pediatric Dentistry
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    • v.29 no.3
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    • pp.337-344
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    • 2002
  • Runx2, a Runt-related osteoblast-specific transcription factor, is essential for osteoblast differentiation and function. Runx2 was identified as a key regulator of osteoblast-specific gene expression through its binding to the OSE2 element present in these genes. However, little is known about the signaling mechanism regulating Runx2 activity. This study examines the role of protein kinase C (PKC) pathway and mitogen-activated protein kinase (MAPK) pathway in regulating Runx2 and bone marker genes (osteopontin; OP, osteocalcin; OC). Luciferase assay and Northern blot analysis suggested that the stimulation of PKC by PMA increased transcription activity of Runx2 and bone marker genes (OP and OC) and also increased expression of Runx2. The stimulation of MAPK by okadaic acid increased transcription activity of Runx2 and bone marker genes (OP and OC). Pretreatment with PD98059 (Erk pathway inhibitor) and SB203580 (P38 pathway inhibitor) prior to PMA treatment decreased PMA stimulated Runx2 activity. Together these results indicate that both PKC and MAPKs are involved in the regulation of Runx2 activity and also the stimulation of Runx2 transcriptional activity by the PKC pathway is through activation of MAPK pathway.

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[$Ca^{2+}$ Sensitization Mechanism in Stretch-induced Myogenic Tone

  • Kim, Jung-Sup;Ryu, Sung-Kyung;Ahn, Duck-Sun;Kang, Bok-Soon;Lee, Young-Ho
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.1
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    • pp.33-39
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    • 2002
  • It has been suggested that $Ca^{2+}$ sensitization mechanisms might contribute to myogenic tone, however, specific mechanisms have not yet been fully identified. Therefore, we investigated the role of protein kinase C (PKC)- or RhoA-induced $Ca^{2+}$ sensitization in myogenic tone of the rabbit basilar vessel. Myogenic tone was developed by stretch of rabbit basilar artery. Fura-2 $Ca^{2+}$ signals, contractile responses, PKC immunoblots, translocation of PKC and RhoA, and phosphorylation of myosin light chains were measured. Stretch of the resting vessel evoked a myogenic contraction and an increase in the intracellular $Ca^{2+}$ concentration $([Ca^{2+}]_i)$ only in the presence of extracellular $Ca^{2+}$. Stretch evoked greater contraction than high $K^+$ at a given $[Ca^{2+}]_i.$ The stretch-induced increase in $[Ca^{2+}]_i$ and contractile force were inhibited by treatment of the tissue with nifedipine, a blocker of voltage-dependent $Ca^{2+}$ channel, but not with gadolinium, a blocker of stretch-activated cation channels. The PKC inhibitors, H-7 and calphostin C, and a RhoA-activated protein kinase (ROK) inhibitor, Y-27632, inhibited the stretch-induced myogenic tone without changing $[Ca^{2+}]_i.$ Immunoblotting using isoform-specific antibodies showed the presence of $PKC_{\alpha}$ and $PKC_{\varepsilon}$ in the rabbit basilar artery. $PKC_{\alpha},$ but not $PKC_{\varepsilon},$ and RhoA were translocated from the cytosol to the cell membrane by stretch. Phosphorylation of the myosin light chains was increased by stretch and the increased phosphorylation was blocked by treatment of the tissue with H-7 and Y-27632, respectively. Our results are consistent with important roles for PKC and RhoA in the generation of myogenic tone. Furthermore, enhanced phosphorylation of the myosin light chains by activation of $PKC_{\alpha}$ and/or RhoA may be key mechanisms for the $Ca^{2+}$ sensitization associated with myogenic tone in basilar vessels.