• 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.

• Bulletin of the Korean Chemical Society
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• v.24 no.12
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• pp.1799-1804
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• 2003

Acetolatate synthase(ALS) catalyzes the first common step in the biosynthesis of valine, leucine, isoleucine in plants and microorganisms. ALS is the target of several classes of herbicides, including the sulfonylureas, the imidazolinones, and the triazolopyrimidines. To elucidate the roles of arginine residues in tobacco ALS, chemical modification and site-directed mutagenesis were performed. Recombinant tobacco ALS was expressed in E. coli and purified to homogeneity. The ALS was inactivated by arginine specific reagents, phenylglyoxal and 2,3-butanedione. The rate of inactivation was a function of the concentration of modifier. The inactivation by butanedione was enhanced by borate, and the inactivation was reversible on removal of excess butanedione and borate. The substrate pyruvate and competitive inhibitors fluoropyruvate and phenylpyruvate protected the enzyme against inactivation by both modifiers. The mutation of well-conserved Arg198 of the ALS by Gln abolished the enzymatic activity as well as the binding affinity for cofactor FAD. However, the mutation of R198K did not affect significantly the binding of FAD to the enzyme. Taken together, the results imply that Arg198 is essential for the catalytic activity of the ALS and involved in the binding of FAD, and that the positive charge of the Arg is crucial for the interaction with negatively charged FAD.

• Journal of Plant Biotechnology
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• v.36 no.4
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• pp.397-402
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• 2009

Ornithinine carbamoyltransferase (OTC) is an enzyme that catalyzes the key step in arginine biosynthesis in bacteria and plants. OTC is also involved in the urea cycle and deficiency of the enzyme in human leads to disease. The argF gene encoding OTC has been reported in many bacteria and few plants. Here we report the characterization of a gene encoding OTC from rice (OsOTC). Analysis of a cDNA sequence from rice revealed that the full-length open reading frame of OsOTC consisted of 367 amino acids, corresponding to a protein of approximately 39.7 kDa. The predicted amino acid sequence of OsOTC harbor distinct five OTC signature sites and is highly homologous to that of enzymes of plants, animals and many bacterial OTCs. Expression of OsOTC in argF mutants of Escherichia coli showed that the gene was able to functionally complement to the mutant. These results suggest that the OsOTC encode a protein for ornithine carbamoyltransferase in rice.

• Journal of Plant Biology
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• v.36 no.1
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• pp.1-8
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• 1993

Polyamine contents and the activities of their main biosynthetic enzymes, arginine decarboxylase (ADC) and ornithine decarboxylase(ODC), were investigated in Populus leaf segments during adventitious shoot regeneration with the addition of 1 millimolar polyamine synthesis inhibitors. From the study of polyamine synthesis inhibitors, ODC was found to be the principal route of polyamine biosynthesis in adventitious shoot regeneration of Populus leaf segments. The ADC inhibitor difluoromethyl arginine(DFMA) and ODC inhibitor difluoromethyl ornithine(DFMO) strongly reduced the putrescine content. On the contarary, DCHA, an inhibitor of spermidine synthase, increased it. Spermidine content was decreased with the treatment of each polyamine synthesis inhibitor, but the inhibitory effect of DCHA was stronger than any other polyamine inhibitor. The decreased polyamine level by polyamine synthesis inhibitors was restored with the exogenously applied polyamine. Comparing the polyamine contents with the adventitious shoot regeneration rate, were observed a close correlation between spermidine content and adventitious shoot regeneration of Populus leaf segments.

• YAKHAK HOEJI
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• v.38 no.1
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• pp.24-30
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• 1994

Nitric oxide(NO) synthase was identified and characterized by determining the L-citrulline formed in the NO-Arg pathway in pancreatic tissues. NO synthase activities in chicken pancreas were dependent upon the concentration of L-Arg which is the substrate molecule for the NO synthase, the amount of the enzyme protein used, and linearly on the incubation time. NO synthase in mouse pancreas was found to be constitutive, not induced by lipopolysaccharide treatment. In vitro NO synthase activities of chicken pancreas were inhibited 36%, 21%, 12% and 44% by $200\;{\mu}M$ of MMA, DMA, D'MA and NAME respectively. These results suggest the presence of NO and NO synthase in the pancreas.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.14-20
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• 1991

- The purpose of this work is to investigate the effects of various substrates on biosynthesis of ethylene by the Kudzu strain of Pseudomonas syn'ngae pv. Phaseolicola causing halo blight. In the intact cell of P. sym'ngue, optimal condition for ethylene production was achieved at p1-I 7.5 and $30^{\circ}C$ for 9 to 10 hours of culture. Ethylene was most effectively produced from amino acids such as Asn, Gln, Asp ans Glu, compared to those of various kinds of sugars. While ethylene production from $\alpha$-ketoglutarate ($\alpha$-KG) was gradually increased throughout 51 hours incubation period tested. Ethylene production derived from citrate, $\alpha$-KG and oxalacetate as well as a few amino acids was further enhanced by the addition of histidine or arginine. In cell-free ethylene-forming system, ethylene was most effectively produced from $\alpha$-KG, compared to those from citrate, oxalacetate, Glu, Arg, or Asp, at 0.5 mM among the range from 0.25 mM to 5 mM. Anlinooxyacetate, an inhibitor of a pyridoxal phosphate-linked enzyme, completely inhibited ethylene evolution derived from Glu but not affect that derived from $\alpha$-KG. The results obtained in this work suggest that $\alpha$-KG might be a direct precursor of ethylene production in this organism than any other substrates tested.

• Journal of Microbiology
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• v.34 no.4
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• pp.355-362
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• 1996

Complementation cloning of the argC, E, B, D, F, and G genes in Corynebacterium glutamicum was done by transforming the genomic DNA library into the corresponding arginine auxotrophs fo Escherichia coli. Recombinant plasmids containing 6.7 kb and 4.8kb fragments complementing the E. coli argB mutant were also able to complement the E. coli argC, E, A, D, and F mutants, indicating the clustered organization of the arginine biosynthetic genes within the cloned DNA fragments. The insert DNA fragments in the recombinant plasmids, named pRB1 AND pRB2, were physically mapped with several restriction enzymes. By further subcloning the entire DNA fragment containing the functions and by complementation analysis, we located the arg genes in the order of ACEBDF on the restriction map. We also determined the DNA nucleotide sequence of the fragment and report here the sequence of the argB gene. When compared to that with the mutant strain, higher enzyme activity of N-acetylglutamate kinase was detected in the extract of the mutant carrying the plasmid containing the putative argB gene, indicating that the plasmid contains a functional argB gene. Deduced amino acid sequence of the argB gene shows 45%, 38%, and 25% identity to that from Bacillus strearothermophilus, Bacillus substilus, and E. coli respectively. Our long term goal is genetically engineering C. glutamicum which produces more arginine than a wild type strain does.

• YAKHAK HOEJI
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• v.38 no.2
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• pp.149-157
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• 1994

The role of nitric oxide(NO) as neurotransmitter in non-adrenergic non-cholinergic (NANC) relaxation induced by electrical stimulation has been studied in circular muscle strips of the rabbit gastric fundus. In the presence of atropine and guanethidine, low frequency$(1{\sim}20\;Hz)$ and short trains (5s) of electrical stimulation induced the frequency-dependent relaxations which were not affected by adrenergic and cholinergic blockage, but abolished by tetrodotoxin, a nerve conductance blocker. L-NNA, a stereospecific inhibitor of NO biosynthesis, inhibited the relaxations induced by electrical stimulation but not affected the relaxation to exogenous NO. The effect of L-NNA was prevented by L-arginine, the precursor of the NO biosynthesis, but not by its enantiomer, D-arginine. Exogenous administration of NO$(10{\sim}100\;{\mu}M)$ caused the concentration-dependent relaxation which showed a similarity to those obtained with electrical stimulation. Hemoglobin, a NO scavenger, abolished the NO-induced relaxations and also markedly inhibited those evoked by electrical stimulation. Application of adenosine triphosphate$(1{\sim}10\;{\mu}M)$ induced concentration-independent contractions, but in high dose caused temporary contraction followed by relaxation which was not affected by L-NNA. Exogenous vasoactive intestinal polypeptide$(10{\sim}100\;nM)$ induced the concentration-dependent relaxation, while its effects were slower in onset and more persistent than those induced by short trains and low frequencies of electrical stimulation. Based on above results, it is suggested that NO is the principal neurotransmitter of NANC nerve at relaxation induced by short trains and low frequencies of electrical stimulation in the rabbit gastric fundus.

• Korean Journal of Microbiology
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• v.25 no.1
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• pp.46-51
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• 1987

In order to increase the production of tryptophan by maximizing expression of recombinant trp plasmid, Klebsiella pneumoniae KC 105(pheA tyrA trpE trpR tyrR) was genetically modified. KC 107, inosine monophospate(IMP) auxotroph from KC 105 and KC 108, histidine(His) auxotroph from KC 107 were also derived respectively to increase phosphoribosylpyrophosphate(PRPP) production which is required for tryptophan biosynthesis. From KC 107 phosphoribosylpyrophosphate consumption which is required for tryptophan biosynthesis. From KC 107 and KC 108, KC 109 and KC 110, both arginine auxotrophs were derived respectively. To investigate the expression of recombinant trp plasmid in the selected K. pneumoniae mutants, the auxotrophic mutants were transformed with recombinant trp plasmids pSC 101-$trpE^{FBR}$, pSC 101-trpL(.DELTA.att) $trpE^{FBR}$ (pSC 101-trp-AF). Amount of tryptophan produced and activities of tryptophan synthase of $trpR^{-}$ mutant (KC 100) and $tyrR^{-}$ mutnat(KC 105) containing recombinant plasmid pSC 101-trp operon were increased by 30-40% as compared with KC 99(pheA tyrA trpE) containing recombinant plasmid pSC 101-trp operon. Activities of tryptophan synthase and production of tryptophan of KC 108 ($His^{-}$) and KC 109($Arg^{-}$) containing recombinant plasmid pSC 101-trp operon were increase by two-fold as compared with KC 107 containing pSC 101-trp operon.

• Journal of Plant Biology
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• v.31 no.3
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• pp.239-247
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• 1988

Spermidine, spermine and IAA promoted an increase in $\beta$-1, 4-endoglucanase activity in hypocotyls of Glycine max. The optimal concentration for the increase of the enzyme activity was 10-6 M for spermidine, 10-8 M for spermine and 10-6 M for IAA. However, IAA had innocuous effect on arginine decarboxylase and ornithine decarboxylase activites, and the content of polyamine. Such cumulative results suggest that the increase in $\beta$-1, 4-endoglucanase activity by IAA is not attributed by the effect on the biosynthesis of polyamine by IAA but spermidine, spermine and IAA induce cell wall loosening and therefore extension growth of cells.