• Title/Summary/Keyword: Metalloenzyme

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THE CHANGE OF EXTRACELLUAR ALKALINE PHOSPHATASE EXPRESSION IN HYPERTROPHIC SCAR IN RABBITS (가토의 비후성 반흔에서 세포외 알칼리성 인산효소 발현의 변화)

  • Cho, Young-Ki;Ryu, Sun-Yul
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.21 no.1
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    • pp.23-28
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    • 1999
  • ALP (alkaline phosphatase) is a membrane-bound metalloenzyme that is expressed in osteoblasts, hepatocytes, lung, kidney, endothelial cells, leukocytes and other cells. Normal soft tissue and skin show little tissue nonspecific ALP (TN-AP), However, scar tissue contains high levels of TN-AP activity, and in fact, TN-AP is expressed intensely in regenerating connective tissue after the wounding. The purpose of this study was to evaluate the change of ALP expression in hypertrophic scar model in rabbits and the effect of triamcinonolone on ALP expression. Adult male New Zealand white rabbits, weighing about 2.5 kg, were used. After full-thickeness wounding over the ventral surface of each ear, either saline (control ear) or triamcinolone (contralateral ear) was injected on day 16. Rabbits were sacrificed on day 3, 7, 15, 17, 19, 23, and the specimens were retrieved en bloc. Histologic and immunohistochemical examinations of tissue samples were done. The results obtained were as follows: On day 3, ALP reaction was observed on fibroblasts and inflammatory cells in wound margin. On day 7, ALP reaction was more intense than day p in capillaries, inflammtory cells, and fibroblasts behind newly formed epithelium. On day 15, ALP reaction was lessened in both groups and appeared mainly in subepidermal capillary network, Since day 17, ALP reaction was lessened in both groups and weaker in triamcinolone-injected group than in saline-injected group. These results suggest that ALP reaction isn't increased in triamcinolone-injected scar and triamcinolone reduces scar not by increasing TN-AP expression but other mechanism.

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Presence of Carbonic Anhydrase III-like Protein in Shaggy Sea Raven, Hemitripterus villosus (삼세기(Shaggy sea raven, Hemitripterus villosus)의 carbonic anhydrase III에 관한 연구)

  • Kweon, Rok Eun;Kho, Kang Hee
    • Journal of Life Science
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    • v.24 no.2
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    • pp.186-190
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    • 2014
  • Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

Characteristics and Action Pattern of Alkaline Protease from Streptomyces gviseus HC-1141 (Streptomyces griseus HC-1141이 생성하는 Alkaline Protease의 특성 및 작용양상)

  • Choi, Cheong;Chung, Yung-Gun;Sung, Sam-Kyung;Choi, Kwang-Soo;Lee, Jae-Sung;Cho, Young-Je;Chun, Sung-Sook
    • Microbiology and Biotechnology Letters
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    • v.20 no.3
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    • pp.295-301
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    • 1992
  • An alkaline protease producing microorganism was isolated from soil and identified as Streptomyces griseus HC-1141. The optimum pH and temperature for the purified enzyme activity were 8.0 and $60^{\circ}C$, respectively. The enzyme was relatively stable in the pH range of 7.0-9.0 and at the temperature below $60^{\circ}C$. The activity of purified enzyme was inhibited by $Hg^{2+}$, $Cu^{2+}$, $Zn^{2+}$, $Ba^{2+}$ and $Fe^{2+}$, whereas activated by $Mn^{2+}$ and $Ca^{2+}$. $\varepsilon$-Amino caproic acid, 2,4-dinitrophenol and iodine did not show inhibitory effect on the activity of alkaline protease, but p-chloromercuribenzoic acid, ethylendiaminetetraacetic acid showed inhibitory effect on the enzyme activity. These result suggested that the protease was metalloenzyme, and require a reactive SH group for the activity. The reaction of this enzyme follows typical Michaelis-Menten kinetics with the $K_m$ value of $2.229{\times}10^{-4}$M and the $V_{max}$ of $46.08 {\mu}$g/min for casein. The activation energy for the alkaline protease calculated by Arrhenius equation was 3.643 kcal/mol. This enzyme hydrolyzed casein more rapidly than the hemoglobin and egg albumin.

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Studies on Effect of Dietary Zine on Tissue Trace Elements in the Rat (식이아연(食餌亞鉛)이 흰쥐의 조직중(組織中) 미양금속(微量金屬)에 미치는 영향(影響))

  • Suk, Young-Gun
    • Journal of Nutrition and Health
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    • v.5 no.2
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    • pp.91-103
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    • 1972
  • Zinc is one of the essential trace elements in the living organism for growth and health. The first identified metalloenzyme, carbonic anhydrase, is a zinc compound and several others have been described since. Among zinc deficiency syndromes in animals porcine parakeratosis has been successfully treated with zinc supplements, and in man a syndrome of anemia, hypogonadism, hepatosplenomegaly, and dwarfism, prevalent in parts of Iran and Egypt, has been ascribed to lack of zinc in the diet. Dietary zinc excess in the rat is manifested by a hypochromic, microcytic anemia, poor growth, reduction in liver catalase and cytochrome oxidase. The present study is an attempt to delineate the changes of tissue contents of trace elements, especially of iron, copper and zinc in liver and kidneys of the rats. Weanling albino rats, weighing 60 to 80gm. were used in this experiments. The rats were housed in cages with aluminum floors and received feed and distilled water ad libitum. Animals were divided into three groups, control, low zinc diet and high zinc diet groups. The high zinc diet group was subdivided into 0.5% Zn and 0.7%Zn groups. The supplementary copper or iron was added to the high dietary zinc groups. The animals were sacrificed and the tissues were washed several times with deionized water. The wet digested samples were analyzed by Hitachi Model 207 atomic absorption spectro-photometer for the determination of iron, copper and zinc in the liver and kidney. Hemoglobin level in the blood was measured by cyanmethemoglobin method. The results of this study are as follows: 1) All rats fed high zinc diets and low zinc diets gained less weight than control. Weight gain was not improved by the supplementary copper or iron and both. 2) Hemoglobin concentration was decreased significantly in the rats fed high zinc diets and less in the low zinc diet. Supplementary copper and iron to the higher zinc diet appeared to give some improvement of anemia. 3) The iron contents of the liver and kidneys were significantly decreased in the high zinc groups and the reduction was more significantly in the rats receiving higher zinc diet (0.7%). The supplementary copper caused a further depression of liver iron. On the other hand, the iron, added to the high zinc diet lessoned the severity of the decrease in liver iron and caused kidney iron to be maintained almost at the level found in the rats fed by zinc and supplementary copper diet. 4) High zinc diets did not change copper content of the liver and kidney. Supplementary copper elevated the concentration in the liver and kidney and added iron had no effect on the accumulation of copper in the liver and kidneys. 5) The high zinc diets caused marked increases of zinc content in the liver and kidney. Supplementary iron to the high zinc diet caused increases of zinc contents of liver and kidneys.

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Expressional Analysis of Superoxide Dismutase in Olive Flounder (Paralichthys olivaceus) against Viral Hemorrhagic Septicemia Virus Infection (Viral hemorrhagic septicemia virus (VHSV) 감염에 대한 넙치 superoxide dismutase(Of-SOD)의 발현분석)

  • Lee, Young Mee;Kim, Jung-Eun;Noh, Jae Koo;Kim, Hyun Chul;Park, Choul-Ji;Park, Jong-Won;Kim, Kyung-Kil;Lee, Jeong-Ho
    • Journal of Life Science
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    • v.24 no.12
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    • pp.1371-1377
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    • 2014
  • Superoxide dismutase is a family of important antioxidant metalloenzymes and catalyzes the dismutation of toxic superoxide anions into dioxygen and hydrogen peroxide. A recent study identified the partial superoxide dismutase (SOD) gene in olive flounder (Paralichthys olivaceus). The same study reported that it strongly induced benzo[a]pyrene and that it was an indicator of aquatic oxidative stress responses. However, its transcriptional response against viral infection has not been investigated. In the present study, the spatial and temporal expression profiles were analyzed to investigate the function of Of-SOD in the antiviral response. The Of-SOD transcripts were ubiquitously detected at various levels in diverse tissues in a real-time PCR. The expression of Of-SOD was significantly higher in the muscles, liver, and brain but extremely low in the stomach and spleen. Following a VHSV challenge, the expression of Of-SOD increased within 3 h in the kidneys and decreased to the original level 2 days postchallenge. In muscle, liver, and brain, Of-SOD mRNA was similarly up-regulated at 3-6 h postchallenge and then decreased to the basal level. Although the expression pattern and induction time differed slightly depending on the tissue, the transcript of Of-SOD consistently increased in the acute infection response, but the expression was low in the chronic response. The expression of Of-SOD was induced after the VHSV infection, and Of-SOD was probably involved in the immune response against the viral challenge. These results suggest that SOD may play important roles in the immune defense system of P. olivaceus and perhaps contribute to the protective effects against oxidative stress in olive flounder.