• Title/Summary/Keyword: $\gamma$-Glutamyl transpeptidase

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A Spectrophotometric Assay for ${\gamma}$-Glutamyl Transpeptidase Activity

  • Hwang, Se-Young
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.360-363
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    • 1994
  • A colorimetric assay for $\gamma$-glutamyl transpeptidase ($\gamma$-CTP; E.C 2.3.2.2) employing 2, 4, 6-trinitrobenzene sulfonate (TNBS) to detect the amount of disappeared acceptor via transpeptidation, has been developed. Under the experimental conditions using L-$\gamma$-glutamyl ethyl ester and L-phenylalanine as $\gamma$-glutamyl donor and acceptor, respectively, it was found that the decreased absorbance of yellow color at 420 nm was strictly related to the amount of L-$\gamma$-glutamyl-L-phenylalanine (L-$\gamma$-Glu-L-Phe) formed, which was determined by DEAE-cellu-lose column chromatography. Concentrations of the enzyme and $\gamma$-glutamyl products were able to be determinedin the nanogram and nanomoles per milliliter range, respectively, with high precision and reliability. This novel assay system may therefore be a useful means for understanding of catalytic function of the $\gamma$-CTP spectrophotometrically without any usage of sophisticated instruments.

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Effects of Bromate on the Glutathione Synthesis in Various Organs of Rats (Bromate가 흰쥐의 장기 Glutathione 함량에 미치는 영향)

  • 김나영;강혜옥;이무강;최종원
    • Journal of Life Science
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    • v.13 no.5
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    • pp.626-633
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    • 2003
  • The effects of bromate administration on glutathione were studied in rats. The contents of glutathione in the liver and kidney were significantly decreased but the alteration was not significant in lung and blood by bromate adminstration. The decrease occurred without concomitant increases in oxidized glutathione (GSSG) or in the GSSG/GSH+GSSG ratio. The activities of $\gamma-glutamyl$ cysteine synthetase in the liver and kidney were decreased by bromate administration. $\gamma-Glutamyl$ transpeptidase activities was significantly decreased in the kidney and not significantly decreased in the lung of bromate treated-rats. These results suggest that the decreased synthesis of glutathione by bromate may be an important reason for the decreased level of glutathione in the liver and kidney, thus the decreased glutathione transport would be a factor on the changes of glutathione contents in bromate-treated rats.

Purification and Properties of $\gamma$-Glutamyl Transpeptidase from Bacillus sp. KUN-17

  • Hwang, Se-Young;Ryang, Jun-Hwan;Lim, Wang-Jin;Yoo, Ick-Dong;Kunio Oishi
    • Journal of Microbiology and Biotechnology
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    • v.6 no.4
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    • pp.238-244
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    • 1996
  • $\gamma$-Glutamyl transpeptidase ($\gamma$-GTP; EC 2.3.2.2) present in the culture filtrate of Bacillus sp. KUN-17 was purified 400-fold through a consecutive procedure including organic precipitation and column chromatography. The enzyme has an estimated molecular weight of 70, 000 and consists of hetero-subunits with molecular weights of 42, 000 and 22, 000. In vitro optimal conditions for those transfer and hydrolysis reactions appeared to be pH 7.0 at $50^{\circ}C$ and pH 8.4 at $40^{\circ}C$, respectively. The denatured enzyme recovered most of its $\gamma$-GTP activity by removing detergents such as sodium dodecyl sulfate (SDS) or urea with dialysis. The enzyme showed higher affinities against a number of amino acids as $\gamma$-glutamyl acceptors than glycylglycine in the following order: L-valine, L-methionine, L-glutamic acid or L-as-paragine, L-alanine. Also, it was shown that L-glutamine was the most suitable $\gamma$-glutamyl donor for the transfer reaction among those tested. Amino acids generally inhibited the enzyme activity for the transfer reaction, but not for the hydrolysis reaction.

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AN EXPERIMENT OF ${\gamma}-GLUTAMYL$ TRANSPEPTIDASE ON PERIODONTAL INFLAMMATION (치주염증시 ${\gamma}-Glutamyl$ transpeptidase의 연구)

  • Lee, Seok-Cho;Lim, Jong-Deuk;Yoo, Kang-Rgeol;Oh, Kwi-Ok;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.23 no.3
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    • pp.517-525
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    • 1993
  • Glutathione(GSH),a tripeptide thiol, found in virtually all cells, functions in metabolism tranasport and cellular protection. It protects cells against the destructive effects of reactive oxygen intermediates and free radicals. Also ${\gamma}-Glutamyl$ transpeptidase(${\gamma}-GTT$), an enzyme of major importance in GSH metabolism, initiates GSH degradation. In order to explore the $GSH-{\gamma}-GTT$ system as periodontal disease activity indicator, we observed the ${\gamma}-GTT$ and arachidonic acid metabolits according to clinical groups(Control, Adult periodontitis, Rapidly progressive Periodontitis). From the experiments, the following results were obtained. 1. When compared with normal, ${\gamma}-GTT$ of A. P. and R. P. P. were increased, and only the change of ${\gamma}-GTT$ of R. P. P. was statistically significant(P<0. 05). 2. The amounts of arachidonic acid metabolites were not different with statistical significance among the clinical groups. 3. ${\gamma}-GTT$ may by useful adjuncts as new cytoprotective indicator and periodontal disease activity indicator in accordance with positive corelation pocket depth, attachment level and ${\gamma}-GTT$.

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An Essential Histidine Residue in the Catalytic Mechanism of the Rat Kidney γ-Glutamyl Transpeptidase

  • Kim, Soo-Ja;Ko, Moon-Kyu;Chai, Kyu-Yun;Cho, Seong-Wan;Lee, Woo-Yiel
    • Bulletin of the Korean Chemical Society
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    • v.28 no.2
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    • pp.271-275
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    • 2007
  • γ -Glutamyl transpeptidase (EC 2.3.2.2) plays a key role in glutathione metabolism by catalyzing the transfer of the γ -glutamyl residue and hydrolysis of glutathione. The functional residues at the active site of the rat kidney γ -glutamyl transpeptidase were investigated by kinetic studies at various pH, the treatment of diethylpyrocarbonate (DEPC), and photooxidation in presence of methylene blue. An ionizable group affecting the enzymatic activity with an apparent pKa value of 7.1, which is in the range of pKa values for a histidine residue in protein, was obtained by examining the pH-dependence of kinetic parameters. The pH effect on the photoinduced inactivation rate of the enzyme corresponds to that expected for the photooxidation of the free histidine. The involvement of a histidine in the catalytic site of the enzyme was further supported by DEPC modification accompanied by an increase in absorbance at 240 nm, indicating the formation of Ncarbethoxyhistidine. The histidine located at the position of 382 in the precursor of the enzyme is primarily suspected based on the amino acid sequence alignment of the transpeptidases from various organisms.

Protective Effect of Biological Osmolytes against Heat- and Chaotropic Agent-Induced Denaturation of Bacillus licheniformis γ-Glutamyl Transpeptidase

  • Lo, Huei-Fen;Chi, Meng-Chun;Lin, Min-Guan;Lan, Yuan-Gin;Wang, Tzu-Fan;Lin, Long-Liu
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1457-1466
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    • 2018
  • In the present study, the stabilizing effect of four different biological osmolytes on Bacillus licheniformis ${\gamma}$-glutamyl transpeptidase (BlGGT) was investigated. BlGGT appeared to be stable under temperatures below $40^{\circ}C$, but the enzyme retained less than 10% of its activity at $60^{\circ}C$. The tested osmolytes exhibited different degrees of effectiveness against temperature inactivation of BlGGT, and sucrose was found to be the most effective among these. The use of circular dichroism spectroscopy for studying the secondary structure of BlGGT revealed that the temperature-induced conformational change of the protein molecule could be prevented by the osmolytes. Consistently, the molecular structure of the enzyme was essentially conserved by the osmolytes at elevated temperatures as monitored by fluorescence spectroscopy. Sucrose was further observed to counteract guanidine hydrochloride (GdnHCl)-and urea-induced denaturation of BlGGT. Taken together, we observed evidently that some well-known biological osmolytes, especially sucrose, make a dominant contribution to the structural stabilization of BlGTT.

The Schizosaccharomyces pombe Gene Encoding ${gamma}-Glutamyl$ Transpeptidase I Is Regulated by Non-fermentable Carbon Sources and Nitrogen Starvation

  • Kim, Hong-Gyun;Park, Hey-Jung;Kang, Hyun-Jung;Lim, Hye-Won;Kim, Kyung-Hoon;Park, Eun-Hee;Ahn, Ki-Sup;Lim, Chang-Jin
    • Journal of Microbiology
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    • v.43 no.1
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    • pp.44-48
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    • 2005
  • In our previous study, the first structural gene (GGTI) encoding ${\gamma}-glutamyl$ transpeptidase was cloned and characterized from the fission yeast Schizosaccharomyces pombe, and its transcription, using the GGTI-lacZ fusion gene, containing the 1,085 bp upstream region from the translational initiation point, was found to be enhanced by sodium nitroprusside and L-buthionine-(S,R)-sulfoximine (BSO). In the present work, regulation of the GGTI gene was further elucidated. Non-fermentable carbon sources, such as acetate and ethanol, markedly enhanced the synthesis of ${beta}-galactosidase$ from the GGTI-lacZ fusion gene. However, its induction by non-fermentable carbon sources appeared to be independent of the presence of the Pap1 protein. Nitrogen starvation also gave rise to induction of GGTI gene expression in a Pap1-independent manner. The three additional fusion plasmids, carrying 754, 421 and 156 bp regions, were constructed. The sequence responsible for the induction by non-fermentable carbon sources and nitrogen starvation was identified to exist within a -421 bp region of the GGTI gene. Taken together, the S. pombe GGTI gene is regulated by non-fermentable carbon sources and nitrogen starvation.

Changes of Serum Gamma-Glutamyl Transpeptidase Values in Experimentally Induced Toxic Hepatitis, Obstructive Jaundice and Pancreatitis in Dogs (실험적으로 일으킨 개의 중독성(中毒性) 간염(肝炎), 폐쇄성(閉鎖性) 황달(黃疸) 및 췌장염(膵臟炎)에 있어서 혈청(血淸) Gamma-Glutamyl Transpeptidase치(値)의 변화)

  • Seung, Won-Pyo;Choi, Hee-In
    • Korean Journal of Veterinary Research
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    • v.23 no.1
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    • pp.25-35
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    • 1983
  • In order to assess the diagnostic aid of serum gammaglutamyl transpeptidase values in hepatitis, obstructive jaundice and pancreatitis, four groups of 14 health dogs were subjected to the gastric intubatin of $CCl_4$(1.5ml/kg of body weight), the ligation of common bile duct, the ligation of pancreatic ducts and the injection of chloroform(0.2ml/kg of body weight) in the parenchyma of the pancreas. Some serum enzymes serum glutamic pyruvic transaminase(SGPT), serum glutamic oxalacetic transaminase(SGOT), total bilirubin, amylase and lipase known to be indicative of hepatic and pancreatic diseases were monitored. In comparision of these enzymes, gamma-glutamyl transpeptidase(GGTP) valuers were determined in these dogs before and after the experimental procedures. The results were summarized as follows: 1. In $CCl_4$ intoxication gorup, there were no significant changes in serum GGTP activities(mean: 6.0~14.6 IU/L). 2. In bile duct ligation group, serum GGTP activities shelved marked increases, beginning at postsurgical day 1 and rose the highest mean value(342.7 IU/L) on day 12. Then the activities never approached to the base-line values. 3. After the ligation of pancreatic ducts and the injection of chloroform in the pancreas, serum GGTP activities did not rise throughout the experiment. 4. SGPT:GGTP ratio did not increase in bile duct ligation group, but increase markedly in $ccl_4$ intoxication group. 5. The results indicated that serum GGTP values or SGPT:GGTP ratio could provide valuable indicators for differential diagnosis between hepatobiliary obstruction and hepatocellular disease.

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The health effects of low blood lead level in oxidative stress as a marker, serum gamma-glutamyl transpeptidase level, in male steelworkers

  • Su-Yeon Lee;Yong-Jin Lee;Young-Sun Min;Eun-Chul Jang;Soon-Chan Kwon;Inho Lee
    • Annals of Occupational and Environmental Medicine
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    • v.34
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    • pp.34.1-34.13
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    • 2022
  • Background: This study aimed to investigate the association between lead exposure and serum gamma-glutamyl transpeptidase (γGT) levels as an oxidative stress marker in male steelworkers. Methods: Data were collected during the annual health examination of workers in 2020. A total of 1,654 steelworkers were selected, and the variables for adjustment included the workers' general characteristics, lifestyle, and occupational characteristics. The association between the blood lead level (BLL) and serum γGT level was investigated by multiple linear and logistic regression analyses. The BLL and serum γGT values that were transformed into natural logarithms were used in multiple linear regression analysis, and the tertile of BLL was used in logistic regression analysis. Results: The geometric mean of the participants' BLLs and serum γGT level was 1.36 ㎍/dL and 27.72 IU/L, respectively. Their BLLs differed depending on age, body mass index (BMI), smoking status, drinking status, shift work, and working period, while their serum γGT levels differed depending on age, BMI, smoking status, drinking status, physical activity, and working period. In multiple linear regression analysis, the difference in models 1, 2, and 3 was significant, obtaining 0.326, 0.176, and 0.172 (all: p < 0.001), respectively. In the multiple linear regression analysis stratified according to drinking status, BMI, and age, BLLs were positively associated with serum γGT levels. Regarding the logistic regression analysis, the odds ratio of the third BLL tertile in models 1, 2, and 3 (for having an elevated serum γGT level within the first tertile reference) was 2.74, 1.83, and 1.81, respectively. Conclusions: BLL was positively associated with serum γGT levels in male steelworkers even at low lead concentrations (< 5 ㎍/dL).

Evaluation of the Herbal Extract Mixture for the Effects of Hair-Regrowth Compared to 3% Minoxidil; Elongation of Anagen Period on C3H Mice (모발생장기 유도 C3H 생쥐에 있어서 미녹시딜과 생약추출 혼합 조성물의 모발 재성장 유도 효능)

  • 이계호;한선일;박길흥;권영이
    • YAKHAK HOEJI
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    • v.47 no.1
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    • pp.14-19
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    • 2003
  • The hair cycle consists of three phases, growth (anagen), involution (catagen) and quiescence (telogen) phases. In order to evaluate hair re-growth effect of herbal extracts mixture containing the 70% ethanol extracts of Polygoni Multiflori Radix, Mori Cortex Radicis, Gingko Biloba Folium and Pine bud, we have examined the induction of the anagen phase and/or elongation of the anagen period using C3H mice. Morphological examination was done by Hattori' and Ogawa's method. Enzyme activities of ${\gamma}$-glutamyl transpeptidase (${\gamma}$-GT) and alkaline phosphatase (ALP) was detected by Bessey-Lovry-Brock's method. Enzyme activity as a biochemical marker of hair cycle was investigated in the third hair cycle period of C3H mice after depilation. 3% Minoxidil treated group and herbal extract mixture treated group were shown 3 days earlier initiation of anagen than control group. In cycling mouse skin, ${\gamma}$-GT activity is pronounced during anagen and greatly diminished during telogen. Herbal extract mixture has shown promising hair re-growth effect on hair follicular cycles of C3H mice.