• Title/Summary/Keyword: isozymes

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Distributional Patterns of Phospholipase C Isozymes in Heart and Brain of Spontaneously Hypertensive and Normotensive Rats

  • Choi, Ji-Woong;Cho, Young-Jin;Cha, Seok-Ho;Lee, Kweon-Haeng;Lee, Sang-Bok
    • The Korean Journal of Physiology and Pharmacology
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    • v.1 no.4
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    • pp.385-392
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    • 1997
  • The phospholipase C (PLC)-mediated intracellular signal transduction pathway is considered to be involved in the regulation of blood pressure. However, little information is available concerning the distributional and functional significance of PLC in the genetic hypertensive rats. As the first step of knowing the role of PLC on hypertension, we investigated the distribution of 6 PLC isozymes $(PLC-{\beta}1,\;-{\beta}3,\;-{\beta}4,\;-{\gamma}1,\;-{\gamma}2\;and\;-{\delta}1)$ in the heart and brain, which are concerned with hypertension, in the normotensive Wistar-Kyoto rat (WKY) and spontaneously hypertensive rat (SHR) using the western blotting and immunocytochemistry. The immunoreactivities of PLC isozymes in brain were detected, but there were no distributional and quantitative differences between the WKY and SHR. In the heart, but the immunoreactivities to $PLC-{\beta}1$ and $-{\gamma}2$ in the SHR were higher than those in WKY. In immunocytochemistry to confirm these western blotting data, $PLC-{\beta}1$ and $-{\gamma}2$ were localized in cardiac myocytes and the intensities of immunoreactivity in SHR were stronger than that in WKY. These results suggest that $PLC-{\beta}1$ and $-{\gamma}2$ would have possibility to concern with the establishment of spontaneous hypertension.

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Serum Enzyme Values after Extracorporeal Circulation (체외순환후의 혈청효소치 변동에 관한 연구)

  • Lee, Sang-Ho;Kim, Chong-Whan
    • Journal of Chest Surgery
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    • v.14 no.2
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    • pp.144-152
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    • 1981
  • The effect of cardiopulmonary bypass procedures on the blood components were studied through the measurements of several serum enzymes, SGOT, SGPT, CPK and LDH with its isozymes in the patients who had surgery for their heart diseases. They wre 85 patients: 40 Males and 45 Females. Their ages ranged widely between one and fifty-six. They were divided into 5 groups: Group I.patent ductus arteriosus without extracorporeal circulation, Group II-atrial septal defect, Group III-ventricular septal defect, Group IV-tetralogy of Fallot, and Group V-valve replacement. Generally serum enzymes revealed the increased values after surgery and the tendency returning toward preoperative levels. With the high total serum LDH levels seen uniformly after operation which persisted long in patients with replaced valves, the changes of LDH-isozymes were further analyzed. The isozymes, especially LDI and LDs increased their activities postoperatively, and the other fractions varied little. And $LD_1$/$LD_2$ ratios also showed similar rises and falls while the elevated values lasted longer in the patients with cardiac valve replacement. The significant values of LDH-isozymes in predicting the hemolysis occuring after extracorporeal circulation and in following the patients whose valves were replaced have been further discussed with the special stress placed on the $LD_1$/$LD_2$ratios.

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Over-Expression of Phospholipase D Isozymes Down-Regulates Protein Kinase CKII Activity via Proteasome-Dependent CKIIβ Degradation in NIH3T3 Cells

  • Yoon, Soo-Hyun;Min, Do Sik;Bae, Young-Seuk
    • Molecules and Cells
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    • v.27 no.3
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    • pp.299-305
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    • 2009
  • Over-expression of phospholipase D (PLD) 1 or PLD2 down-regulated CKII activity in NIH3T3 cells. The same results were found with catalytically inactive mutants of PLD isozymes, indicating that the catalytic activity of PLD is not required for PLD-mediated CKII inhibition. Consistent with this, 1-butanol did not alter CKII activity. The reduction in CKII activity in PLD-over-expressing NIH3T3 cells was due to reduced protein level, but not mRNA level, of the $CKII{\beta}$ subunit. This PLD-induced $CKII{\beta}$ degradation was mediated by ubiquitin-proteasome machinery, but MAP kinase and mTOR were not involved in $CKII{\beta}$ degradation. PLD isozymes interacted with the $CKII{\beta}$ subunit. Immunocytochemical staining revealed that PLD and $CKII{\beta}$ colocalize in the cytoplasm of NIH3T3 cells, especially in the perinuclear region. PLD binding to $CKII{\beta}$ inhibited $CKII{\beta}$ autophosphorylation, which is known to be important for $CKII{\beta}$ stability. In summary, the current data indicate that PLD isozymes can down-regulate CKII activity through the acceleration of $CKII{\beta}$ degradation by ubiquitin-proteasome machinery.

Separation and Purification of Soybean $\beta$-amylase Isozymes (대두 $\beta$-amylase Isozyme의 분리 및 정제)

  • 지의상
    • The Korean Journal of Food And Nutrition
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    • v.3 no.2
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    • pp.149-160
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    • 1990
  • The soybean $\beta$-amylase ($\alpha$-1, 4-glucan maltohydrolase, EC 3.2.1.2) is composed of seven isozymes(I', I, II, III, IV, V and VI), and isozyme II and IV are the main components among these. The Purification of $\beta$-amylase isozymes from soybean whey were performed by ammonium sulfate fractionation, CM-Sephadex C-50 column chromatography, DEAE-Sephadex chromatography and Gel filtration. The resulted purity of $\beta$-amylase was throughly confirmed by electrophoresis, and then determined its isoelectric point and molecular weight. The results obtained were as follows, 1. Five active fractions of soybean p-amylase were derived on CM-Sephadex C-50 column chromatography. 2. Seven active bands of p-amylase isozymes were detected by isoelectric focusing gel electrophoresis, and their isoelectric points(I' to VI) were 5.07, 5.15, 5.25, 5.40, 5.55, 5.70 and 5.93, respectively. 3. Isozyme II and IV were main components of soybean $\beta$-amylase. 4. The molecular weights of both isozyme II and IV were determined to be 56,000 daltons by the result of SDS polyacrylamide gel electrophoresis. 5. Km values of main isozyme II & IV for amylopectin were determined to be 2.25 mg/ml, which suggest the same function of each isozyme.

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Selective Homologous Expression of Recombinant Manganese Peroxidase Isozyme of Salt-Tolerant White-Rot Fungus Phlebia sp. MG-60, and Its Salt-Tolerance and Thermostability

  • Kamei, Ichiro;Tomitaka, Nana;Motoda, Taichi;Yamasaki, Yumi
    • Journal of Microbiology and Biotechnology
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    • v.32 no.2
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    • pp.248-255
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    • 2022
  • Phlebia sp. MG-60 is the salt-tolerant, white-rot fungus which was isolated from a mangrove forest. This fungus expresses three kinds of manganese peroxidase (MGMnP) isozymes, MGMnP1, MGMnP2 and MGMnP3 in low nitrogen medium (LNM) or LNM containing NaCl. To date, there have been no reports on the biochemical salt-tolerance of these MnP isozymes due to the difficulty of purification. In present study, we established forced expression transformants of these three types of MnP isozymes. In addition, the fact that this fungus hardly produces native MnP in a high-nitrogen medium (HNM) was used to perform isozyme-selective expression and simple purification in HNM. The resulting MGMnPs showed high tolerance for NaCl compared with the MnP of Phanerochaete chrysosporium. It was worth noting that high concentration of NaCl (over 200 mM to 1200 mM) can enhance the activity of MGMnP1. Additionally, MGMnP1 showed relatively high thermo tolerance compared with other isozymes. MGMnPs may have evolved to adapt to chloride-rich environments, mangrove forest.

Studies on the amino acid metabolism of young rice root (Part 4) -Properties of glutamic oxalacetic transaminase isozymes of rice root- (수도근(水稻根)의 Amino 산(酸) 대사(代謝)에 관(關)한 연구(硏究) -제(第) 4 보(報) 수도근(水稻根)의 Glutamic Oxalacetic Transaminase Isozyme의 성질(性質))

  • Kim, Kwang Sik;Kim, Young Ung
    • Korean Journal of Soil Science and Fertilizer
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    • v.7 no.4
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    • pp.209-213
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    • 1974
  • The isozymes of glutamic oxalacetic transaminase from young rice root were isolated by DEAE-cellulose column chromatography, and investigated some of their enzymic properties. Obtained results were summarized as follows: 1. 93 percent of GOT activity in rice root existed in supernatant fraction, and their specific activity exibited the same pattern. 2. The rice root contained two types of GOT isozyme; cationic and anionic GOT. It seemed that both of them were distributed in supernatant fraction. 3. GOT isozymes of rice root on the pH dependance showed their maximum activity at 7.4. 4. On the stability of GOT isozymes for temperature, anionic GOT was more unstable than cationic GOT. 5. Apparent Michaelis constant for L-aspartate of GOT isozymes from rice root showed 12-14mM in the cationic GOT and 16mM in the anionic GOT, respectively.

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Direct Detection of (1-3)-$\beta$-Glucanase Isozymes in Isoelectrofocusing Gels Using a Dye -Labeled Substrate (염료착색 기질을 이용한 IEF gel에서(1-3)-$\beta$-glucanase 동위효소의 검출)

  • Yun, Song-Joong;Lee, Myong-Chul;Kwon, In-Sook;Kim, Tae-San;Go, Seung-Joo
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.39 no.2
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    • pp.121-127
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    • 1994
  • A procedure for the direct detection of (1-3)-$\beta$-glucanase isozymes in electrophoresis gels was developed. The procedure employed the commercial preparation of AZCL-pachyman as a chromogenic substrate for (1-3)-$\beta$-glucanases. The procedure detected the three basic isozymes which have been known to be expressed in germinating barley kernels. A major acidic and a minor isozymes were also detected in germinating kernels. The procedure was proved to be fast, simple and sensitive enough to be used for the analysis of the expression of (1-3)-$\beta$-glucanase isozymes in plant tissues. The detection limit of the procedure for the commercial preparation of Penicillium (1-3)-$\beta$-glucanase was estimated to be as low as 50$\mu$U. The procedure could be used for the investigation of (1-3)-$\beta$-glucanases in laboratories facilitated with ordinary equipments and research personnel.

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The Functional Role of Phospholipase D Isozymes in Apoptosis (세포사멸에서 Phospholipase D 동위효소의 기능적 역할)

  • Min, Do Sik
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1378-1382
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    • 2014
  • Phospholipase D (PLD) catalyzes the hydrolysis of phospholipid to phosphatidic acid (PA), a lipid secondary messenger. Two forms of PLD isozymes, phosphatidylcholine-specific PLD1 and PLD2, have been identified. PLD has emerged as a critical regulator of cell proliferation and survival signaling, and dysregulation of PLD occurs in a various illnesses, including cancer. PLD activity is essential for cell survival and protection from apoptosis. Overexpression of PLD isozymes or PLD-generated PA attenuates the expression of apoptotic genes and confers resistance to apoptosis. The apoptosis-related molecular mechanisms of PLD remain largely unknown. Recently, the dynamics of PLD turnover during apoptosis have been reported. The cleavage of PLD isozymes as specific substrates of caspase differentially regulates apoptosis. PLD1 is cleaved at one internal site, and PLD2 is cleaved two sites at the front of the N-terminus. The cleavage of PLD1 reduces its enzymatic activity, probably via the dissociation of two catalytic motifs, whereas the cleavage of PLD2 does not affect the catalytic motifs and its activity. Thus, PLD2 maintains antiapoptotic capacity, despite its cleavage. Therefore, the differential cleavage pattern of PLD isozymes by caspase affects its enzymatic activity and antiapoptotic function. Thus, PLD is considered a potential target for cancer therapy. We summarize recent studies regarding the functional role of PLD in apoptosis.

Analysis on Enzymatic Browning in Pine Needles (솔잎에서 발생하는 효소학적 갈변작용에 관한 분석)

  • Kong, Kwang-Hoon;Park, Hee-Joong;Choi, Sang-Sook;Cho, Sung-Hye;Kim, Yong-Tae
    • Analytical Science and Technology
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    • v.12 no.3
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    • pp.256-259
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    • 1999
  • Tyrosinases are related to the enzymatic browning of plants and attract the major scientific interest for the prevention of it. Three tyrosinase isozymes ($P_1$, $P_2$ and $P_3$) from pine needles were purified to homogeneity and characterized the factors that affect their activities. The L-ascorbic acid and ${\beta}$-mercaptoethanol notably inhibited the enzymatic activities of the three isozymes. The sodium diethyldithiocarbamate was a competitive inhibitor of isozymes with the $K_i$ values of $P_1$(0.030 mM), $P_2$(0.015 mM) and $P_3$(0.019 mM), respectively. Their enzyme activities were however, increased by the addition of most metal ions. The optimum pH for the three isozymes was 9.0~9.5 and the optimum temperatures ranged from 55 to $60^{\circ}C$ using L-DOPA as substrate.

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In vitro inhibitory effects of Yangguksanhwa-tang and Taeumjowi-tang on CYP450 isozymes (양격산화탕과 태음조위탕 추출물의 in vitro CYP450 효소 활성 억제 효과)

  • Ha, Hye-Kyung;Jin, Seong-Eun;Shin, Hyeun-Kyoo
    • Journal of Society of Preventive Korean Medicine
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    • v.16 no.3
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    • pp.129-137
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    • 2012
  • Objective : Herb-drug interactions have become an important issue because of the consumption of herbal remedies has increased in the world. Yangguksanhaw-tang (Liang ge san huo-tang) and Taeumjowi-tang (Tai yin tiao wei-tang) are typical herbal formulas on Sasang constitution medicine (four-constitution medicine). This study was aimed at evaluating the effects of Yangguksanhaw-tang and Taeumjowi-tang on drug metabolizing enzymes, cytochrome P450 (CYP450) isozymes. Methods : Vivid$^{(R)}$ CYP450 Screening Kits were used to measure of CYP3A4, CYP2C19, CYP2D6 and CYP2E1 activities. This method is based on the use of fluorescent CYP450 substrates that are efficiently metabolized by specific CYP450 isozymes to yield a product with altered fluorescent properties. The percent inhibitions of CYP450s by herbal formulas were calculated. Results : Yangguksanhaw-tang inhibited CYP2C19 and CYP2E1 activities higher than that other CYP450 isozymes. The $IC_{50}$ values of CYP2C19 and CYP2E1 were 159.83 ${\mu}g/mL$ and 261.40 ${\mu}g/mL$, respectively. The CYP2E1 activity was inhibited ($IC_{50}=215.17{\mu}g/mL$) higher than that other CYP450 isozymes by Taeumjowi-tang. Conclusions : These results suggest that Yangguksanhaw-tang may inhibit the metabolism of co-administered drugs whose primary route of metabolism is via CYP2C19 or CYP2E1. Taeumjowi-tang could inhibit the metabolism of co-administered drugs, which are substrates for CYP2E1. Therefore, co-administration of the herbal formulas and other conventional drugs should be undertaken with care.