• Title/Summary/Keyword: Guanosine nucleotide

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The Origin of Ribityl Side Chain of Riboflavin in Ashbya gossypii (Ashbua gossypii에서의 리보플라빈 측쇄의 기원)

  • 최원자;임정빈
    • Korean Journal of Microbiology
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    • v.23 no.3
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    • pp.167-171
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    • 1985
  • In order to investigate the origin of the ribityl froup of riboflavin and the involvement of GTP cyclohydrolase II in the riboflavin pathway, we studied the incorporation of $^{14}C-labeled$ guanosine using a well known riboflavin over producer, Ashbya gossypii.Cells were grown in a media containing $(U- ^{14}C)$ guanosine and the riboflavin and GMP were isolated and purifired by column chromatography. The isolated compounds, riboflavin and GMP were labeled in the ribityl and ribosyl side chain and the isoalloxazine and guannine moiety. By comparing the specific radioactivity of each compound we reached a conclusion that the ribose of guanosine is converted directly to the rivityl moiety of riboflavin. The results indicate that biosynthesis of the vitamin begins at the level of a guanosine compound and also suppory the involvement of GTP cyclohydrolase II in one of the early steps in the biosynthetic pathway.

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Allosteric Properties of Hafnia alvei Aspartase by Nucleotide Effectors

  • Noh, Hak-Joon;Kwon, Si-Joong;Kim, Ki-Tae;Lee, Chang-Hyun;Yoon, Moon-Young
    • BMB Reports
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    • v.33 no.5
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    • pp.366-369
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    • 2000
  • The nucleotide effects of Hafnia alvei aspartase were investigated. Purine nucleosides, such as adenosine and guanosine, increased the aspartase activities; whereas, purine nucleotides, such as AMP, ATP, GTP and IMP, caused little change in the aspartase activities. However, pyrimidine derivatives, such as cytidine and CTP, decreased the aspartase activity. The nucleotide and nucleoside effects by the limited trypsin-treated aspartase were similar to those of a native enzyme. These results indicate that the COOH-terminal region and an allosteric site might be located away from each other. The initial velocity study in the presence of adenosine showed that $K_m$ for aspartate was decreased to one-sixth of that in the absence of adenosine, but $V_{max}$ was unchanged. The significance of the distinct allosteric effect for the enzyme-nucleotide interaction is discussed.

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Purification and Characterization of Pyrimidine Nucleotide N-Ribosidase from Pseudomonas oleovorans

  • YU, Tae-Shick
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.573-578
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    • 2005
  • Pyrimidine nucleotide N-ribosidase (pyrimidine 5'-nucleotide phosphoribo(deoxyribo)hydrolase/pyrimidine 5'-nucleotide nucleosidase, EC 3.2.2.10) catalyzes the breakdown of pyrimidine 5'-nucleotide into pyrimidine base and ribose(deoxyribo)-5-phosphate. However, detailed characteristics of the enzyme have not yet been reported. The enzyme was purified to homogeneity 327.9-fold with an overall yield of $6.1\%$ from Pseudomonas oleovorans ATCC 8062. The enzyme catalyzed cytidine monophosphate (CMP) and uridine monophosphate (UMP), but not adenosine monophosphate (AMP) and guanosine monophosphate (GMP). The enzyme optimally metabolized CMP at pH 6.0 and UMP at around 8.5, and the optimum temperature for the overall enzyme reaction was found to be $37^{\circ}C$. The $K_m$ values of the enzyme for CMP (at pH 6.0) and UMP (at pH 8.5) were 1.6 mM and 1.1 mM, respectively. AMP, deoxyCMP, and deoxyUMP were very effective inhibitors of the reaction. Double-reciprocal plots obtained in the absence and in the presence of AMP revealed that this inhibitory effect was of the mixed competitive type with respect to the breakdown of CMP and of the noncompetitive type with respect to the breakdown of UMP. In the presence of AMP, the enzyme followed sigmoid kinetics with respect to each substrate.

Ras GTPases and Ras GTPase Activating Proteins (RasGAPs) in Human Disease (Ras GTPase 및 Ras GTPase activating protein과 사람의 질병)

  • Chang, Jong-Soo
    • Journal of Life Science
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    • v.28 no.9
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    • pp.1100-1117
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    • 2018
  • The Ras superfamily of small G-proteins acts as a molecular switch on the intracellular signaling pathway. Upon ligand stimulation, inactive GTPases (Ras-GDP) are activated (Ras-GTP) using guanine nucleotide exchange factor (GEF) and transmit signals to their downstream effectors. Following signal transmission, active Ras-GTP become inactive Ras-GDP and cease signaling. However, the intrinsic GTPase activity of Ras proteins is weak, requiring Ras GTPase-activating protein (RasGAP) to efficiently convert RAS-GTP to Ras-GDP. Since deregulation of the Ras pathway is found in nearly 30% of all human cancers, it might be useful to clarify the structural and physiological roles of Ras GTPases. Recently, RasGAP has emerged as a new class of tumor-suppressor protein and a potential therapeutic target for cancer. Therefore, it is important to clarify the physiological roles of the individual GAPs in human diseases. The first RasGAP discovered was RASA1, also known as p120 RasGAP. RASA1 is widely expressed, independent of cell type and tissue distribution. Subsequently, neurofibromatosis type 1 (NF1) was discovered. The remaining GAPs are affiliated with the GAP1 and synaptic GAP (SynGAP) families. There are more than 170 Ras GTPases and 14 Ras GAP members in the human genome. This review focused on the current understanding of Ras GTPase and RasGAP in human diseases, including cancers.

Nucleotide Analysis in Korean Dairy Products Using High- Performance Liquid Chromatography with Diode Array Detector

  • Won, Jong-Eun;Bang, Han-Yeol;Kwak, Byung-Man;Park, Jong-Su;Kim, Gui-Ran;Kwon, Joong-Ho
    • Food Science of Animal Resources
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    • v.39 no.1
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    • pp.93-101
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    • 2019
  • Nucleotides play important roles in numerous intracellular biochemical processes and are used in infant formulas and other dairy products. However, domestic analytical methods for assessing nucleotide content in products have not yet been established, and therefore, methods for determining nucleotide content are urgently required. A rapid and simple analytical method for determining the content of five types of nucleotides in dairy products was improved using solid phase extraction clean-up and high-performance liquid chromatography with diode array detector. The extraction solvent used in the AOAC method was not well dissolved and was changed to hydrophilic EDTA-Na. In addition, the results obtained using the isocratic elution method and a single wavelength were similar to those obtained using the AOAC method, and the time taken for analysis was shortened from 40 min to 25 min. The process of method validation revealed the following parameters: accuracy (84.69%-102.72%), precision (1.51%-6.82%), linearity (0.999), and limit of detection (cytidine 5'-monophosphate, 0.09 mg/L; uridine 5'-monophosphate, 0.11 mg/L; adenosine 5'-monophosphate, 0.12 mg/L; guanosine 5'-monophosphate, 0.11 mg/L; and inosine 5'-monophosphate, 0.14 mg/L). The method was also used to determine the nucleotide concentration in 25 samples (infant formulas, 1.99-29.39 mg/100 g; and cow milk, 0.28-0.83 mg/100 g). The newly improved method was appropriate for analyzing nucleotides in infant formulas and other dairy products faster when compared to conventional methods.

Studies on the Mechanical Activities of Rabbit Myometrium V. Effects of Acetylcholine, Oxytocin and Prostagla, din F2α on Cyclic Nucleotide Levels of Rabbit Whole Uterus (가토 척출 자궁근의 운동성에 관한 연구 V. Acetylcholine, PGF2α 및 Oxytocin의 자궁 수축기전에 관한 연구)

  • Lee, Chang-Eop;Kwun, Jong-Kuk;Lee, Joong-Sup;Yang, Il-Suk;Lee, Mun-Han
    • Korean Journal of Veterinary Research
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    • v.22 no.1
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    • pp.1-8
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    • 1982
  • The effect of acetylcholine, oxytocin and prostaglandin $F_{2{\alpha}}$ ($PGF_{2{\alpha}}$) on cyclic nucleotide levels in estrogen-primed rabbit whole uterus were studied in the presence and absence of 1-methyl-3-isobutyl xanthine (MIX), a phosphodiestrase inhibitor, and indomethacin, a prostagandin inhibitor. In the absence of MIX, acetylcholine increased guanosine 3', 5'-cyclic monophosphate (cGMP), but had no effect on adenosine 3', 5'-cyclic monophosphate (cAMP) levels. In contrast, oxytocin had no influence on cGMP, but decreased cAMP levels. $PGF_{2{\alpha}}$ increased cGMP and decreased cAMP levels. MIX increased both cAMP and cGMP levels. Oxytocin and $PGF_{2{\alpha}}$ further increased cGMP levels, indicating activation of guanylate cyclase activity. The ratio of cAMP/cGMP was decreased by uterine stinulants both in presence and absence of MIX. Indomethacin elevated cAMP and cGMP revels. The effects of uterine stimulants in the presence of indomethacin on cyclic nucleotide levels were varied from tissue to tisse. In general, oxytocin decreased cGMP and $PGF_{2{\alpha}}$ increased cAMP/cGMP levels, but the effects were statisically nonsignicficant. The cAMP/cGMP ratio was increased by uterine stimulant in the presence of indomethacin. In conclusion, uterine stimulants eased cAMP/cGMP ratio which indicates that the uterine stimulants have opposing effects on adenylate cyclase and guanylate cyclase activities. The endometrium plays a role in the regulation of cyclic nucleotide levels and uterine contraction by means of PG synthesis. Indomethacin has an unknown activities besides both of PG synthetase and phosphodiesterase inhibitions.

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Studies on the Highly-phosphorylated Nucleotides during the Differentiation of Aspergillus niger (검정곰팡이의 분화(分化)에 따르는 균체내(菌體內)의 고인산(高燐酸)뉴크레오티드의 소장(消長)에 관한 연구(硏究))

  • Kim, Jong-Hyup
    • The Korean Journal of Mycology
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    • v.10 no.2
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    • pp.57-65
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    • 1982
  • Highly phosphorylated nucleotides were investigated to assure whether the eucaryotic Aspergillus niger produce these substances or not during the differentiation. Investigation was extended to see how organic phosphate interacts with inorganic polyphosphate during development, and high molecular weight RNA-polyphosphate complex was detected in 2.6% polyacrylamide gel by electrophoresis. Guanosine tetraphosphate was found in vesicle and phialide forming mycelia and spore forming body by PEI cellulose TLC. It is revealed that guanosine tetraphosphate is a common substance for spore formation in eucaryotic microorganisms as well as in procaryotic. Especially, prior to sporulation, protein bound RNA and protein bound phosphate may occur as a result of reorganization of cellular materials. The evidence was obtained by the fact of differential increase of optical density ratio between the samples from different developmental stages of this fungus. In 2.6% polyacrylamide gel which was run to electrophoresis, high molecular weight RNA (mostly rRNA) was found to couple and to make RNA-polyphosphate complex. The complex was examined with enzymes and radioactive isotope of $^{32}P$. (enzymic test was not reported here.) RNA-polyphosphate complex might be another sort of highly phosphorylated nucleotide or rRNA beside guanosine-tetraphosphate.

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Development of natural Seasoning Based on Mushroom (버섯 천연조미료 제조에 관한 연구)

  • 박미자;이종숙;이별나;이재성
    • Journal of the East Asian Society of Dietary Life
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    • v.11 no.3
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    • pp.196-203
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    • 2001
  • This study was conducted to provide basic data for the development of natural seasoning. The quality chatacteristics of water exacts from Pleurotus ostreatus and Lentinus edodes were determind. Also, savory taste, fragrance, color and overall preference was evaluated. The preference test on the seasonings prepared from Pleurotus ostreatus and Lentinus edodes with the addition of sea tangle was also performed. The crude protein content of dried, Lentinus edodes and Pleurotur ostreatus was 20.4% and 30.4% respectively. The total organic acid contents were higher in Plourotus ostreatus than in Lentinus edodes extracts. The total free amino acid adn total nucleotide contents were higher in Pleurotus ostreatus than in Lentinus edodes extracts. Three nucleotide, adenosine monophosphate(AMP) , guanosine monophosphate(GMP), xanthine monophosphate(XMP) were found in mushroom seasonings while inosine monophosphate(IMP) was not detected. Pleurotus ostreatus extract showed higher score in savory taste and overall acceptability than Lentinus edodes extract, Pleurotur ostreatus seasoning added with 1% sea tangle and Lentinus edodes seasoning added with 1.5% sea tangle were rated as high in overall quality In a sensory test of foods cooked with mushroom seasoning. It was shown that panels in their twenties well used to the taste of monosodium glutamate while the people in their thirties preferred the specific flavor of the mushroom seasoning.

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Regulation of a Novel Guanine Nucleotide Binding Protein Tissue Transglutaminase ($G{\alpha}_n$).

  • Im, Mie-Jae
    • BMB Reports
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    • v.34 no.2
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    • pp.95-101
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    • 2001
  • Tissue transglutaminase (TGII, $G{\alpha}h$) belongs to a family of enzymes which catalyze post-translational modification of proteins by forming isopeptides via $Ca^{2+}$-dependent reaction. Although TGII-mediated formation of isopeptides has been implicated to play a role in a variety of cellular processes, the physiological function of TGII remains unclear. In addition to this Tease activity, TGII is a guanosine triphosphatase (GTPase) which binds and hydrolyzes GTP It is now well recognized that the GTPase action of TGII regulates a receptor-mediated transmembrane signaling, functioning as a signal transducer of the receptor. This TGII function signifies that TGII is a new class of GTP-binding regulatory protein (G-protein) that differs from "Classical" heterotrimeric G-proteins. Regulation of enzyme is an important biological process for maintaining cell integrity. This review summarizes the recent development in regulation of TGII that may help for the better understanding of this unique enzyme. Since activation and inactivation of GTPase of TGII are similar to the heterotrimeric G-proteins, the regulation of heterotrimeric G-protein in the transmembrane signaling is also discussed.

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