• Journal of Microbiology and Biotechnology
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• 제4권4호
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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.

• 생명과학회지
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• 제13권5호
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• pp.626-633
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• 2003

백서에 bromate의 장기간 섭취로 간 및 신장의 glutathion량이 감소되는데, 간과 신장은 유사한 감소양상을 나타냈고, 폐 및 혈액에서는 감소하는 경향은 있었으나 통계적인 유의성은 없었다. Bromate 섭취로$\gamma-glutamyl$-cysteine synthetase와 $\gamma-glutamyl$ transpeptidase 활성이 감소되었다. 따라서 간과 신장의 glutathione 감소로 $\gamma-glutamyl$cysteine synthetase 활성이 감소됨으로서 glutathione 합성저하에 의해 나타난 결과로 생각되고, 폐에서 $\gamma-glutamyl$cysteine synthetase 및 $\gamma-glutamyl$- transpep-tidase 활성에는 별다른 영향이 없었다. 혈액에서는$\gamma-glutamyl$cysteine synthetase와 $\gamma-glutamyl$ transpeptidase 활성 감소로 glutathione의 혈액내로 유입과 타장기로 유출이 모두 저하되어 glutathione량의 변화가 없는 것으로 생각된다. Bromate에 의한 장기내 glutathione량 감소는 유리기 소거기능이 저하되어 bromate에 의해서 생성된 유리기 제거가 미흡할 것으로 생각되므로 bromate 독성의 한 요인이 될 것으로 추측된다.

• Journal of Microbiology and Biotechnology
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• 제6권4호
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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.

• Journal of Periodontal and Implant Science
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• 제23권3호
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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 ${\gamma}-GTT$.

• Bulletin of the Korean Chemical Society
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• 제28권2호
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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.

• Journal of Microbiology and Biotechnology
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• 제28권9호
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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.

• Journal of Microbiology
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• 제43권1호
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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.

• 대한수의학회지
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• 제23권1호
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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.

• 약학회지
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• 제47권1호
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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.

• BMB Reports
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• 제38권5호
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• pp.609-618
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• 2005

$\gamma$-Glutamyl transpeptidase (GGT, EC 2.3.2.2.) catalyzes the transfer of the $\gamma$-glutamyl moiety from $\gamma$-glutamyl containing ompounds, notably glutathione (GSH), to acceptor amino acids and peptides. A second gene (GGTII) encoding GGT was previously isolated and characterized from the fission yeast Schizosaccharomyces pombe. In the present work, the GGTII-lacZ fusion gene was constructed and used to study the transcriptional regulation of the S. pombe GGTII gene. The synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene was significantly enhanced by NO-generating SNP and hydrogen peroxide in the wild type yeast cells. The GGTII mRNA level was increased in the wild-type S. pombe cells treated with SNP. However, the induction by SNP was abolished in the Pap1-negative S. pombe cells, implying that the induction by SNP of GGTII is mediated by Pap1. Fermentable carbon sources, such as glucose (at low concentrations), lactose and sucrose, as a sole carbon source, enhanced the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in wild type KP1 cells but not in Pap1-negative cells. Glycerol, a non-fermentable carbon source, was also able to induce the synthesis of $\beta$-galactosidase from the fusion gene, but other non-fermentable carbon sources such as acetate and ethanol were not. Transcriptional induction of the GGTII gene by fermentable carbon sources was also confirmed by increased GGTII mRNA levels in the yeast cells grown with them. Nitrogen starvation was also able to induce the synthesis of $\beta$-galactosidase from the GGTII-lacZ fusion gene in a Pap1-dependent manner. On the basis of the results, it is concluded that the S. pombe GGTII gene is regulated by oxidative and metabolic stress.