• Title/Summary/Keyword: PLC

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Biochemical Characterization of Phospholipase C$\delta$from liver of Mud loach (Misgurnus mizolepis) (미꾸라지 간으로부터 포스포리파아제 C델타 단백질의 생화학적 특성)

  • Seo, Jung-Soo;Lim, Sang-Uk;Kim, Na-Young;Lee, Sang-Hwan;Oh, Hyun-Suk;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.67-80
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    • 2005
  • Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.

Homogeneity of Phospholipase C of Bovine Uterus and Seminal Vesicle Compared with Brain Isozymes (소의 자궁 및 고환에서 Phospholipase C의 분리 및 뇌 Isozyme과의 비교 연구)

  • Kim, Jung-Hye;Rhee, Sue-Goo;Lee, Ki-Yung
    • Journal of Yeungnam Medical Science
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    • v.5 no.2
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    • pp.37-45
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    • 1988
  • Phosphoinositide-specific phospholipase C(PI-PLC) is a second messenger of signal transducer on cell membrane. In the previous study, PLC of bovine brain has been purified three isozymes. In this paper, uterus and seminal vesicle have been purified. Two peaks of PI-PLC activity were resolved when bovine uterus and seminal vesicle proteins were chromatographed on a DEAE and phenyl TSK 5PW HPLC column. Each two peak was compared with PI-PLC I, IT and ill from bovine brain and we got the retension time on HPLC. The peak fractions with PLC activity were tested homogeneity with brain PLC monoclonal antibodies(Mab). Mab-labeled affigels were bounded in the range of 73.8%~97.5% with PLC I, IT and III. Homogeneity of fractions were revealed that DEAE F-1 and phenyl F-1-I were highest level of PLC III in uterus and seminal vesicle and DEAE F-2 and phenyl F-2-I were mixed PLC I and II.

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PLC symbol naming rule for auto generation of Plant model in PLC simulation (PLC 시뮬레이션에서 Plant model 자동 생성을 위한 PLC Symbol 규칙)

  • Park, Hyeong-Tae;Wang, Gi-Nam;Park, Sang-Chul
    • Journal of the Korea Society for Simulation
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    • v.17 no.4
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    • pp.1-9
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    • 2008
  • Proposed in the paper is an automated procedure to construct a plant model for PLC simulation. Since PLC programs only contain the control logic without the information on the plant model, it is necessary to build the corresponding plant model to perform simulation. Conventionally, a plant model for PLC simulation has been constructed manually, and it requires much efforts as well as the in-depth knowledge of simulation. As a remedy for the problem, we propose an automated procedure to generate a plant model from the symbol table of a PLC program. To do so, we propose a naming rule for PLC symbols so that the symbol names include enough information on the plant model. By analyzing such symbol names, we extract a plant model automatically. The proposed methodology has been implemented, and test runs were made.

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Cellular Localization and Translocation of Duplication and Alternative Splicing Variants of Olive Flounder Phospholipase C-δ1 (넙치 3가지 타입 인지질가수분해효소(PLC-δ1)의 세포 내 위치 및 이동)

  • Kim, Na Young;Kim, Moo-Sang;Jung, Sung Hee;Kim, Myoung Sug;Cho, Mi Young;Chung, oon Ki;Ahn, Sang Jung
    • Journal of Life Science
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    • v.27 no.11
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    • pp.1369-1375
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    • 2017
  • The purpose of this study was to investigate the cellular characterization of phospholipase C-${\delta}1$ in olive flounders (Paralichthys olivaceus). In general, phospholipase C signaling pathways are distributed in nuclei at plasma membranes and in cytoplasms, although the pathways' nuclear localization mechanisms are unclear. P. olivaceus duplicates type-A PoPLC-${\delta}1$ (PoPLC-${\delta}1A$), which has a high similarity to the human isoform PLC-${\delta}$; type-B PoPLC-${\delta}1$ (PoPLC-${\delta}1B$ [Sf]), which has a low similarity to the human isoform PLC-${\delta}$ and the alternative splice variant PoPLC-${\delta}1B$ (Lf), which has a nuclear localization signal (NLS) and a nuclear export signal (NES) for nuclear imports and exports, respectively. This study confirmed the effects of the cellular localization and translocation of GFP-tagged PoPLC-${\delta}1A$, PoPLC-${\delta}1B$ (Sf) and PoPLC-${\delta}1B$ (Lf). It administered treatments of $Ca^{2+}$ ionophore ionomycin and endoplasmic reticulum (ER)-$Ca^{2+}$ pump inhibitor thapsigargin to hirame natural-embryo (HINAE) cells. A laser-scanning confocal microscope was used. GFP-tagged PoPLC-${\delta}1A$ was distributed to the cellular organelles, rather than to the cytoplasms and cytomembranes, when PoPLC-${\delta}1B$ (Lf) and PoPLC-${\delta}1B$ (Sf) were localized at the plasma membranes. The treatments of ionomycin and thapsigargin showed the accumulation of PoPLC-${\delta}1A$ in the nuclei when PoPLC-${\delta}1B$ (Lf) nucleocytoplasmic shuttling and PoPLC-${\delta}1B$ (Sf) nucleocytoplasmic shuttling were not observed. The results were the first evidence that PoPLC-${\delta}1A$, which contains functional, intact NES sequences, has a main role in nucleocytoplasmic shuttling and translocation in fish.

A Study on Development of Fault diagnosis system for PLC self-diagnostics and its external devices (PLC 자체 고장진단과 그의 외부 소자의 고장 진단 시스템 개발에 관한 연구)

  • Bur, Yone-Gi;Blen, Zeung-Nam
    • Proceedings of the KIEE Conference
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    • 1996.07b
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    • pp.1189-1192
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    • 1996
  • In this paper, a fault diagnosis method is proposed for self-diagnostics of PLC(Programmable Logic Controller), process controller in industrial fields, and diagnosis of its external devices such as sensors and actuators. The aim of this research is proposition of systematic method of fault diagnosis of PLC control system and development of its equipment. A PLC fault diagnosis algorithm consists of self-diagnostics given by PLC makers, Inpuot/Output tracking method by analyzing sequence PLC programs, searching method of past fault cases in database using an expert system, and diagnosis of PLC units such as CPU, DI, and DO board. Finally usability of PLC fault diagnostic system is verified by testing a MELSEC PLC.

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Production of a Phospholipase C by Bacillus cereus and Its Characterization (Bacillus cereus에 의한 C형 인지질 분해효소의 생산과 특성 고찰)

  • 서국화;임용식;이종일
    • KSBB Journal
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    • v.19 no.4
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    • pp.250-256
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    • 2004
  • In this work we have cultivated several B. cereus strains in a complex LB medium in order to study the production of phospholipase C (PLC), and among them B. cereus 318 showed the highest productivity of PLC. Some components, i.e., 5 g/L glucose, 5 g/L yeast extract, 5 g/L peptone, 0.5∼1.0 g/L K$_2$HPO$_4$, 0.02∼0.04 g/L ZnSO$_4$$.$7H$_2$O and 3 g/L NaHCO$_3$ were found to be optimal for the high production of PLC by B. cereus 318. Optimal culture temperature and pH were found to be 30$^{\circ}C$ and pH 7.5 for the PLC production, respectively. Optimum reaction temperature and pH of the PLC produced by B. cereus 11 and 318 were 45$^{\circ}C$ and pH 4.0, while they were 50$^{\circ}C$ and pH 7.0 for the PLC by B. cereus 559. The PLC produced by B. cereus was activated by Mn$\^$2+/, Co$\^$2+/ and dimethyl sulfoxide (DMSO), but its activity was inhibited by Cu$\^$2+/ and partially by glycerol, isopropanol and sodium dodecyl sulfate (SDS).

Product Life Cycle in view of Market Risk Management (Market Risk Management 관점에서 본 Product Life Cycle)

  • Shin, On-Myung;Kim, Young-Ei
    • Journal of Distribution Science
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    • v.7 no.1
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    • pp.91-104
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    • 2009
  • The Purpose of this study is to reveal interactive relation between Product Life Cycle and Market Risk Management. PLC analyzes consumer buying behavior, volatility of price and sales at market place from the viewpoint of Marketing so that company can improve management result. This study is attempt to analyze PLC from a comprehensive and integrating angle by using MRM. In order to find out relationship between two theories, this study is an extraction of the factors that affects the management result commonly on PLC and MRM. Then, this study analyzes the factors of PLC from the viewpoint of MRM. The result shows that the factors of PLC and MRM have relation with in terms of volatility of price, trade risk and market share.

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Decreased Expression of Phospholipase C-$\beta$2 in Human Platelets with Impaired Function

  • Lee, Sang-Bong;A. Koneti Rao;Lee, Kweon-Haeng;Xu Yang;Bae, Yun-Soo
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.11a
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    • pp.75-84
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    • 1996
  • Platelets from a patient with a mild inherited bleeding disorder and abnormal platelet aggregation and secretion show reduced generation of inositol 1,4,5-trisphosphate (IP$_3$), mobilization of intracellular Ca$\^$2+/, and phosphorylation of pleckstrin in response to several G protein mediated agonists, suggesting a possible defect at the level of phospholipase C (PLC) activation. A procedure was developed that allows quantitation of platelet PLC isozymes. After fractionation of platelet extracts by high-performance liquid chromatography, seven, out often known PLC isoforms were detected by immunoblot analysis. The amount of these isoforms in normal platelets decreased in the order PLC-${\gamma}$2 > PLC-${\beta}$2 > PLC-${\beta}$3 > PLC-${\beta}$l > PLC-${\gamma}$ > PLC-$\delta$1 > PLC-${\beta}$4. Compared with normal platelets, platelets from the patient contained approximately one-third the amount of PLC-${\beta}$2, whereas PLC-${\beta}$4 was increased threefold. These results suggest that the impaired platelet function in the patient in response to multiple G protein mediated agonists is attributable to a deficiency of PLC-${\beta}$2. They document for the first time a specific PLC isozyme deficiency in human platelets and provide an unique opportunity to understand the role of different PLC isozymes in normal platelet function.

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Immunohistochemical localization of PLC in rat brain after chronic ECS

  • Hey suk Ihm;You, Je-Kyung;Ryu, Jae-Ryun;Shin, Chan-Young;Ko, Kwang-Ho
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1998.11a
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    • pp.197-197
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    • 1998
  • Chronic electroconvulsive shock(ECS) was shown to Increase phosphatidylinositol-4,5-bisphosphate(PIP$_2$) breakdown and the activity of PLC with the accumulation of inositol-1,4,5-triphosphate(IP3). The purpose of the present study was to determine the effect of ECS on the expression of phospholipase C(PLC) isotypes in rat brain. Two groups of animals were prepared: sham and ECS treated groups. Rats in ECS treated groups received maximal ECS(70mA, 0.5second, 60㎐) by constant current stimulator through ear-clip to induce tonic extension seizures for 12 consecutive days. The expression of PLC isotypes in rat brain was determined by immunohistochemical procedure using sagital section of rat brain. The immunoreactivity of PLC${\beta}$1 was observed in corpus striatum, hippocampus, thalamus and that of PLC${\gamma}$1 in corpus striatum, hippocampus, thalamus, frontal cortex, parietooccipital cortex, limbic forebrain, pons, medulla, superior colliculus, inferior colliculus, rest of midbrain. The amount of PLC was analyzed by Western blot using antibodies against PLC${\beta}$1 and PLC${\gamma}$1. Chronic ECS reduced the immunoreactivity of PLC${\beta}$1 in corpus striatum, hippocampus, thalamus but had little effect on PLC${\gamma}$1. To quantify this change, quantitative Western blot using antibodies against PLC${\beta}$1 and PLC${\gamma}$1 was conducted. The immunoreactivity of PLC${\beta}$1 in ECS treated rat whole brain was decreased by 40 % in cytosolic fraction and 26 % in membrane fraction. This different effect of ECS on PLC isotypes may results from the difference of their activation mechanisms and the different effects of ECS on them. The results from the present study suggest that chronic ECS primalily affects neurotransmitter receptors related IP$_3$ signaling in rat brain.

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A Study of PLC Simulation for Automobile Panel AS/RS (자동차 패널 자동창고 시스템의 PLC 시뮬레이션 적용 연구)

  • Ko, Min-Suk;Koo, Lock-Jo;Kwak, Jong-Geun;Hong, Sang-Hyun;Wang, Gi-Nam;Park, Sang-Chul
    • Journal of the Korea Society for Simulation
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    • v.18 no.3
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    • pp.1-11
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    • 2009
  • This paper illustrates a case study of PLC logic simulation in a car manufacturing system. It was developed to simulate and verify PLC control program for automobile panel AS/RS. Because car models become varied, the complexity of supply problem is increasing in the car manufacturing system. To cope with this problem, companies use the AS (automated storage) and RS (retrieval system) but it has logical complexity. Industrial automated process uses PLC code to control the AS/RS, however control information and control codes (PLC code) are difficult to understand. This paper suggests a PLC simulation environment, using 3D models and PLC code with realistic data. Data used in this simulation is based on realistic 3D model and I/O model, using actual size and PLC signals, respectively. The environment is similar to a real factory; users can verify and test the PLC code using this simulation before the implementation of AS/RS. Proposed simulation environment can be used for test run of AS/RS to reduce implementation time and cost.