• Title/Summary/Keyword: GTP

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Antagonists of Phosphatidylinositol 3-Kinase Block Phosphorylation-Dependent Activation of the Leukocyte NADPH Oxidase in a Cell-Free System

  • Park, Jeen-Woo
    • BMB Reports
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    • v.30 no.3
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    • pp.182-187
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    • 1997
  • The NADPH oxidase of phagocytes catalyzes the reduction of oxygen to $O_2^-$ at the expense of NADPH. The enzyme is dormant in resting neutrophils and becomes activated on stimulation. During activation, $p47^{phox}\;(\underline{ph}agocyte\;\underline{ox}idase\;factor)$, a cytosolic oxidase subunit, becomes extensively phosphorylated at a number of serines located between S303-S379. Oxidase activation can also be achieved by the addition of phosphorylated recombinant $p47^{phox}$ by protein kinase C in the cell-free system in the presence of $GTP{\gamma}S$. The cell-free activation is inhibited by wortmannin and LY294002. specific inhibitors of phosphatidylinositol 3kinase (PI 3-kinasel) These results indicate that PI 3-kinase may playa pivotal role in the activation of NADPH oxidase.

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Ric-8B Modulates the Function of Alpha Subunit of Go

  • Kim, Seung-Hyun;Ghil, Sung-Ho
    • Biomedical Science Letters
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    • v.13 no.2
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    • pp.127-133
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    • 2007
  • Heterotrimeric GTP binding proteins (G proteins) mediate signal generated by neurotransmitter and hormones. Among all G proteins, Go is the most abundant in brain but its role in brain is not clearly understood. To determine the function of the alpha subunit of Go (Go$\alpha$), we search for the interacting partner of Go$\alpha$ in brain using yeast two-hybrid system. A resistant to inhibitor of cholinesterase (Ric-8B) was identified as a Go$\alpha$ interacting protein. We confirmed interaction between Go$\alpha$ and Ric-8b employing in vitro affinity binding assay and showed that the Ric-8b increased the function of Go$\alpha$. Our findings indicate that Ric-8b is possible guanine nucleotide exchange factor for Go$\alpha$.

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Isolation and properties of protease Pi in escherichia coli (대장균 세포내 단백질 분해효소, protease Pi의 정제와 특성)

  • 이영섭;곽태환;임정빈;정진하
    • Korean Journal of Microbiology
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    • v.24 no.2
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    • pp.119-126
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    • 1986
  • A periplasmic endoprotease, named protease Pi, was purified to homogeneity from Escherkchia coli by conventional procedure with insulin as substrate. This enzyme degrades insulin and glucagon to trichloroacetic acid-soluble meterials, but shows little or no hydrolysis of bovine serum albumin, casein or globin. Its molecular weight was 110, 000 when determined by gel filtration on Sephacryl S-300 and was 105, 000 when estimated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate. Thus, it appears to be single polypeptide. This snzyme is metalloprotease, since it is completely inhibited by o-phenanthroline and can be activated by addition of divalent metal cations, such as $Mg^{2+}\;and\;Co^{2+}$. It is destinct from protease Ci, a cytoplasmic insulin degrading enzyme, since protease Pi is localized to the periplasm. Since protease Pi selectively degrades GTP cyclohydrolase I, it appears to play a role in the regulation of pteridine biosynthesis.

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Novel $Ca^{2+}$-ATPase Found in the Human Milk Membrane Fraction

  • Cho, Jin-Kook;Kanno, Choemon
    • 한국유가공학회:학술대회논문집
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    • 1997.05a
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    • pp.23-34
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    • 1997
  • Calcium-stimulated ATPase ($Ca^{2+}$-ATPase) which has optimal pH value at 7.0 was found in the membrane fraction of human milk, and its enzymatic properties were studied. The purified $Ca^{2+}$-ATPase required 0.45 mM Ca ion for maximal activity. Among the nucleosides, $Ca^{2+}$-ATPase showed a higher substrate specificity to ATP and UTP than to CTP and GTP. $Ca^{2+}$-ATPase had apparent Km value of 0.065, and V max of 7.63 mol ATP hydrolyzed/mg pro-tein per min, respectively. $Ca^{2+}$-ATPase was potently inhibited by lanthanide, vanadate, and p-chloromercuribenzoate, and inactivated by EDTA, and CDTA and EGTA, but were unaffected by N-ethylmaleimide, $NaN_3$, ouabain, or oligomycin, and was completely inactivated by heating at $60^{\circ}C$ for 10 min. This enzyme activity was concentrated in the membrane fraction of the cream and skim milk membrane, but not founded in bovine milk.

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Purification and characterization of glutamine phosphoribosylpyrophosphate amidotransferase from streptomyces tubercidicus (Streptomyces tubercidicus에서 glutamine phosphoribosylpyrophosphate amidotransferase의 정제 및 특성)

  • 하영칠;유진철
    • Korean Journal of Microbiology
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    • v.29 no.2
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    • pp.97-103
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    • 1991
  • Glutamine phosphoribosylpyrophosphate amidotransferase of Streptomyces tubercidicus was purified and characterized. Molecular weight of the isolated enzyme was determined to be approximately 230,000 and was composed foru identical subunits having a molecular weight of 58,000. This enzyme was strongly inhibited by AMP while considerably inhibited by ATP and GTP. Inhibition effect of enzyme activity by AMP was antagonized by increased concentration of substrate, PRPP, and metal ion (especially, $Mg^{++}$) was essential in both catalytic activity and nucleotide inhibition of this enzyme. Therefore, it was confirmed that end product inhibition of glutamine phosphoribosylpyrophosphate amidotransferase by adenine participated in the regulation of tubercidin biosynthesis from Streptomyces tubercidicus.s.

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Performance Analysis of GGSN IP Buffer Management Strategies in A GPRS Network (GPRS망에서 GGSN IP Buffer 관리기법의 성능분석)

  • 박선춘;마경민;김영진;이형우;조충호
    • Proceedings of the Korean Information Science Society Conference
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    • 2000.10c
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    • pp.334-336
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    • 2000
  • 인터넷의 폭발적인 증가와 다양한 무선 서비스에 대한 계속적인 수요 증가에 따라 용량이 크고, 데이터 전송 속도가 빠르며, 멀티미디어를 지원할 수 있는 무선 서비스가 필요하게 되었다. 비동기식 제3세대 이동통신시스템에서 단말과 인터넷망을 연결하는 GPRS network에서 GGSN의 인터넷 패킷 트래픽을 효과적으로 처리할 수 있는 방안이 연구되어져야 한다. GGSN은 기존의 라우터처럼 하드웨어적으로 구현되는 것이 이상적이지만 GTP (GPRS Tunneling Protocol)가 추가되므로 좀더 복잡한 형태의 gateway가 초기단계에서는 소프트웨어적으로 구현되어질 것이다. 이러한 GGSN의 성능과 용량을 최적화시키기 위한 패킷 트래픽 처리방안에 대한 연구가 필요하다. 본 논문에는 GGSN의 다운 링크상에서 입력되는 각 클래스별 트래픽 모델을 정립하고 트래픽에 대한 특성과 버퍼 관리에 대해 살펴본다.

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Genetically Modified Human Embryonic Stem Cells Relieve Symptomatic Motor Behavior in a Rat Model of Parkinson′s Disease

  • 길광수;이영재;김은영;이창현;이훈택;정길생;박세필;임진호
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.74-74
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    • 2003
  • Embryonic stem cells have several characteristics suitable for cell replacement therapy. To investigate a possibility of using human embryonic stem cell (hESC) as a carrier of therapeutic gene(s), hESC (MB03) was co-transfected with cDNAS coding for tyrosine hydroxylase (TH) and GTP cyclohydrolase Ⅰ (GTPCH Ⅰ) and bulk-selected using neomycin and hygromycin-B. Successful transfection was confirmed by western immunoblotting and RT-PCR. The genetically modified hESC (bk-THGC) relieved apomorphine-induced asymmetric motor behavior by approximately 54% when grafted into striatum of 6-OHDA-denervated rat brain. The number of rotation, however, increased up to 176+18% in 6 weeks when sham-grafted compared with number of rotation before graft. Immunohistochemical staining revealed that the grafted hESC survived and expressed TH for at least 6 weeks while the experiment was continued.

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Characterization of a Noncanonical Purine dNTP Pyrophosphatase from Archaeoglobus fulgidus

  • Im Eun-Kyoung;Hong Chang-Hyung;Back Jung-Ho;Han Ye-Sun;Chung Ji-Hyung
    • Journal of Microbiology and Biotechnology
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    • v.16 no.7
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    • pp.1144-1148
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    • 2006
  • DNA can oxidatively be deaminated by ROS, which converts DNA base amino groups to keto groups and can trigger abnormal mutations, resulting in mutagenesis in organisms. In this study, a noncanonical purine dNTP pyrophosphatase (AfPPase) from a hyperthermophilic archaeon Archaeoglobus fulgidus, which hydrolyzes aberrant nucleoside triphosphates, was overexpressed in E. coli, purified, and characterized. The purified AfPPase showed remarkably high activity for XTP and dITP, suggesting that the 6-keto group of these nucleotides is critical for the reactivity. Under optimal reaction conditions, the reaction rate for these substrates was about 120 times that with dGTP. Therefore, AfPPase may play a significant role in DNA repair by hydrolysis of noncanonical nucleotides before they are misincorporated into DNA.

Optimization of Expression Conditions Enhances Production of Sepiapterin, a Precursor for Tetrahydrobiopterin Biosynthesis, in Recombinant Escherichia coli

  • Park, Eun-Hee;Lee, Won-Heong;Jang, Mi-Hee;Kim, Myoung-Dong
    • Journal of Microbiology and Biotechnology
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    • v.25 no.10
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    • pp.1709-1713
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    • 2015
  • Sepiapterin is a precursor for the synthesis of tetrahydrobiopterin (BH4), which is a wellknown cofactor for aromatic amino acid hydroxylation and nitric oxide synthesis in higher mammals. In this study, a recombinant Escherichia coli BL21(DE3) strain harboring cyanobacterial guanosine 5’-triphosphate cyclohydrolase 1 (GCH1) and human 6-pyruvoyltetrahydropterin synthase (PTPS) genes was constructed to produce sepiapterin. The optimum conditions for T7 promoter–driven expression of GCH1 and PTPS were 30℃ and 0.1 mM isopropyl-β-D-thioglucopyranoside (IPTG). The maximum sepiapterin concentration of 88.1 ± 2.4 mg/l was obtained in a batch cultivation of the recombinant E. coli, corresponding to an 18-fold increase in sepiapterin production compared with the control condition (37℃ and 1 mM IPTG).

The Effect of Angelicae gigantis Radix on the Benzo(a)pyrene-induced Hepatotoxicity in Rats (Benzo(a)pyrene에 의해 유도된 간기능 장해에 미치는 당귀의 효과)

  • 윤수홍;조수열;이윤경;하두현
    • Environmental Analysis Health and Toxicology
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    • v.7 no.1_2
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    • pp.45-51
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    • 1992
  • This study was performed to investigate the effect of Angelicae gigantis Radix extract oil the hepatic detoxifying enzyme activities and lipids. Male sprague-dawley rats were administrated the extracts with benzo(a)pyrene, a hepatotoxic agent, inducing liver damages. Results obtained from this study were as follows: 1. The markedly increased enzyme activities (AST, ALT, LDH, ALP, ${\gamma}$-GTP, GSH-Px) in B(a)P induced groups tended to be decreased by the treatment of the Angelicae gigantis Radix extract. 2. Liver GSH content and lipid peroxide activity were decreased by the administration of the extracts. 3. It tended that the curative effects were better than the protective effects of the extracts.

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