• Title/Summary/Keyword: Arginase

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Purification and Characterization of Two Isolectins with Arginase Activity from the Lichen Xanthoria parietina

  • Molina, M. C.;Vicente, C.
    • BMB Reports
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    • v.33 no.4
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    • pp.300-307
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    • 2000
  • Two glycoproteins were purified and biochemically characterized from the lichen X. parietina. Both behaved as enzymes with arginase activity and haemaglutinins. Secreted arginase (SA) contained galactose and glucose in the saccharide moiety and an isoelectric point of 4.54. The algal binding-protein (ABP) had N-acetyl-glucosamine and glucose as glycosidic residues and an isoelectric point of 3.53. Both proteins had the same molecular mass (58.6 kDa) and the same qualitative amino acidic composition. The results allowed us to consider these glycoproteins as isolectins, which have significant physiological roles in the relationship between photobiont and mycobiont of symbiotic association.

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Inhibition of Fat-Storing Cell Proliferation by a Monomeric Arginase Derived from Perfused Rat Liver

  • Kim, Ki-Yong;Choi, In-Pyo;Kim, Soung-Soo
    • BMB Reports
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    • v.33 no.3
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    • pp.213-220
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    • 2000
  • A fulminant hepatitis is associated with massive liver cell necrosis and a high mortality rate. But survivors regenerate a normal liver and do not have chronic liver disease. This clinical course suggests that the acutely injured livers release a factor that allows a recovery from chronic hepatitis or cirrhosis. The objective of this study was to isolate and characterize an anti-fibrotic factor from acutely damaged rat livers. The liver cell necrosis was prepared from rat by warm ischemical perfusion and the perfusates were assessed against the growth inhibition of fat-storing cells (FSC). A liver-derived growth inhibitory factor (LDGIF) was purified from ischemically damaged rat livers by chromatographies on Sephacryl S-300, CM Sepharose, hydroxyapatite, and Superose 12. The LDGIF was isolated with an overall purification of 194-fold and 40% recovery. Although LDGIF was identified as the rat liver arginase by Nterminal sequence analysis, LDGIF exists as a monomer and the purified native arginase has a trimer form. Furthermore, LDGIF has a lower enzyme activity on the hydrolysis of L-arginine and a higher inhibitory effect on proliferation of FSC than the normal rat liver arginase. The catalytic activity of LDGIF is ascribed to the monomeric characteristics of the LDGIF. Therefore, the inhibitory action of LDGIF might not be due to the arginine depletion by the catalytic activity of arginase. In conclusion, the presence of the LDGIF could interpret the clinical course that serious fibrosis is not found in the liver of patients recovering from severe hepatic necrosis due to fulminant hepatitis, suggesting that this LDGIF may provide a novel target for the prevention and treatment of hepatic fibrosis.

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Binding of Lichen Phenolics to Purified Secreted Arginase from the Lichen Evernia prunastri

  • Legaz, Maria-Estrella;Vicente, Carlos;Pedrosa, Mercedes M.
    • BMB Reports
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    • v.34 no.3
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    • pp.194-200
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    • 2001
  • Secreted arginase from Evernia prunastri thallus has been purified 616-fold from the incubation medium. Purified arginase was resolved as only one peak in a capillary electrophoresis with a pI value of 5.35. The protein contained high amounts of acidic amino acids, such as Asx and Glx, and a relatively high quantity of Ser and Gly. The molecular mass of native, purified arginase was estimated as about 26 kDa by SE-HPLC. Substrate saturated kinetic showed a typical Michaelis-Menten relationship with a K_m value of 3.3 mM L-arginine. Atranorin behaved as a mixed activator of the enzyme (apparent $K_m$ = 0.96 mM); whereas evernic and usnic acid were revealed as non competitive inhibitors (apparent $K_m$ values were 3.16 mM and 3.05 mM, respectively). Kinetics of atranorin binding indicated that saturation was reached from 0.18 ${\mu}mol$ of the total atranorin and the occurrence of multiple sites for the ligand. This agrees with a possible aggregation of several enzyme subunits during the interaction process. A value of binding sites of about 12 was obtained. The binding of evernic acid was saturated from 23 nmol of total phenol. The number of binding sites was about 5. The loss of the binding ability of evernic acid could be interpreted as a single negative cooperatively. Usnic acid behaves in a similar way to evernic acid, although the binding saturation occurs at $0.14\;{\mu}moles$ of the ligand. This binding appears to be unspecific, and has 28 usnic acid binding sites to the protein.

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Korean Red Ginseng Water Extract Restores Impaired Endothelial Function by Inhibiting Arginase Activity in Aged Mice

  • Choi, Kwanhoon;Yoon, Jeongyeon;Lim, Hyun Kyo;Ryoo, Sungwoo
    • The Korean Journal of Physiology and Pharmacology
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    • v.18 no.2
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    • pp.95-101
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    • 2014
  • Cardiovascular disease is the prime cause of morbidity and mortality and the population ages that may contribute to increase in the occurrence of cardiovascular disease. Arginase upregulation is associated with impaired endothelial function in aged vascular system and thus may contribute to cardiovascular disease. According to recent research, Korean Red Ginseng water extract (KRGE) may reduce cardiovascular disease risk by improving vascular system health. The purpose of this study was to examine mechanisms contributing to age-related vascular endothelial dysfunction and to determine whether KRGE improves these functions in aged mice. Young ($10{\pm}3$ weeks) and aged ($55{\pm}5$ weeks) male mice (C57BL/6J) were orally administered 0, 10, or 20 mg/mouse/day of KRGE for 4 weeks. Animals were sacrificed and the aortas were removed. Endothelial arginase activity, nitric oxide (NO) generation and reactive oxygen species (ROS) production, endothelial nitric oxide synthase (eNOS) coupling, vascular tension, and plasma peroxynitrite production were measured. KRGE attenuated arginase activity, restored nitric oxide (NO) generation, reduced ROS production, and enhanced eNOS coupling in aged mice. KRGE also improved vascular tension in aged vessels, as indicated by increased acetylcholine-induced vasorelaxation and improved phenylephrine-stimulated vasoconstriction. Furthermore, KRGE prevented plasma peroxynitrite formation in aged mice, indicating reduced lipid peroxidation. These results suggest KRGE exerts vasoprotective effects by inhibiting arginase activity and augmenting NO signaling and may be a useful treatment for age-dependent vascular diseases.

On Utilization of Canavanine and Activity of Canavanase during Germination and Growth of Canavailia lineata (L.) DC (해녀콩의 발아와 생장시 Canavanine의 이용과 Canavanase의 활성에 대하여)

  • 권영명
    • Journal of Plant Biology
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    • v.29 no.2
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    • pp.85-94
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    • 1986
  • Canavanine content of the cotyledons of Canavalia lineata decreased gradually during germination and growth of seedlings but continued to increase in roots and leaves. After abscission of cotyledons, canavanine content of leaves depleted competely. The activity of canavanase could be detected in leaves and roots, but not in cotyledons. High arginase activity was observed in the cotyledons of seeds at the earlyimbibition period. During the growth of seedlings, cotyledonary canavanine appeared to be transported to the growing of seedlings where it could be utilized through nitrogen metabolic pathways. In crude cell-free extracts of leaves, maximum activities of canavanase or arginase appeared in 30mM Tris-HCl buffer (pH 9.0) or 30mM NaHCO3 buffer (pH 10.0), respectively. The activities of these two enzymes differed from each other when treated with Co2+ or Mn2+. These results support the idea that canavanase and arginase might be different enzymes.

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Studies on Enzyme Activities Responsible for Biosynthesis of Putrescine in Ginseng(Panax ginseng C.A. Meyer) (인삼에서 Putrescine 합성에 관련된 효소활성에 관한 연구)

  • 조병구;조영동
    • Journal of Ginseng Research
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    • v.14 no.1
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    • pp.6-9
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    • 1990
  • Specific activities of ADC and ODC from 2-4 year old ones were higher than that from seedlings whereas those activities were not changed significantly from 2 to 4 years. Generally, activity of ADC was predorminant compared to that of ODC. Free arginine content in roots was much higher than that of leaves. And arginase specific activity from roots was higher than that of leaves. Cumulative results suggest that putrscine formation from ornithine in roots may be more effective than leaves and contribute to putrescine biosynthesis to some extract.

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Modulation of L-Arginine-Arginase Metabolic Pathway Enzymes: Immunocytochemistry and mRNA Expression in Peripheral Blood and Tissue Levels in Head and Neck Squamous Cell Carcinomas in North East India

  • Srivastava, Shilpee;Ghosh, Sankar Kumar
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.16
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    • pp.7031-7038
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    • 2015
  • Background: Arginine may play important roles in tumor progression by providing ornithine for polyamine biosynthesis, required for cell growth. The aim of this work was to determine the expression of arginine metabolic pathway enzymes in head and neck squamous cell carcinoma (HNSCC) in northeast India. Materials and Methods: The expressions of arginase isoforms (ARG1 and ARG2), ornithine aminotransferase (OAT) and ornithine decarboxylase (ODC) were examined in fifty paired HNSCC and adjacent non-tumor tissues by immunohistochemistry. Immunocytochemistry, semiquantitative reverse transcription sq-PCR and quantitative real-time qPCR were used to assess protein and mRNA expressions in peripheral blood of fifty HNSCC patients and hundred controls. Results: ARG1 and ODC protein and mRNA were strongly expressed in peripheral blood from HNSCC patients. No ARG2 expression was observed. In vivo, expression of ARG1, ARG2 and ODC was significantly higher in tumor than in non-tumor tissues. Most tumors expressed low levels of OAT, with no difference in tissues or blood, compared to controls. The absolute extent of maximal ARG1 upregulation with qPCR showed 6.23 fold increase in HNSCC. Conclusions: These findings strongly suggest that in HNSCCs, the ARG1 pathway is stimulated leading to the formation of polyamines as indicated by higher ODC expression, which promote tumor growth.

Effect of thymus extract on the activation of cytotoxic and accessory functions of tumor-associated macrophages

  • Shrivastava, Pratima;Singh, Sukh Mahendra;Singh, Nisha
    • Advances in Traditional Medicine
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    • v.4 no.1
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    • pp.9-17
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    • 2004
  • The present investigation was under taken to study whether the tumor-associated macrophages (TAM) of Daltons lymphoma (DL), a spontaneous transplantable murine T cell lymphoma can be activated to tumoricidal state by crude thymus extract. Intraperitoneal administration of thymus extract to DL-bearing mice resulted in activation of TAM with an enhanced IL-1, TNF and antigen presenting ability. It was found that treatment with thymus extract could also enhance the phagocytic and cytotoxic activity of TAM. However, only a marginal increase in arginase activity was observed. Till date to the best of our knowledge the effect of crude thymus extract on the activation of tumor associated macrophages has not been investigated, this study provides a new piece of information in the area of thymus therapy.