• Title/Summary/Keyword: Cyclase

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Activation of Cryptic hop Genes from Streptomyces peucetius ATCC 27952 Involved in Hopanoid Biosynthesis

  • Ghimire, Gopal Prasad;Koirala, Niranjan;Sohng, Jae Kyung
    • Journal of Microbiology and Biotechnology
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    • v.25 no.5
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    • pp.658-661
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    • 2015
  • Genes encoding enzymes with sequence similarity to hopanoids biosynthetic enzymes of other organisms were cloned from the hopanoid (hop) gene cluster of Streptomyces peucetius ATCC 27952 and transformed into Streptomyces venezuelae YJ028. The cloned fragments contained four genes, all transcribed in one direction. These genes encode polypeptides that resemble polyprenyl diphosphate synthase (hopD), squalene-phytoene synthases (hopAB), and squalene-hopene cyclase (hopE). These enzymes are sufficient for the formation of the pentacyclic triterpenoid lipid, hopene. The formation of hopene was verified by gas chromatography/mass spectrometry.

Is Nitric Oxide Involved in Relaxation of Urinary Bladder\ulcorner

  • Chang, Ki-Churl;Chung, Byung-Ha
    • Biomolecules & Therapeutics
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    • v.3 no.1
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    • pp.58-62
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    • 1995
  • We investigated whether nitric oxide (NO) may serve a role in bladder function by immunohistochemical analysis of the distribution of intrinsic NADPH-diaphorase and functional study of isometric tension recordings via a photo-induced adequate nitric oxide (PIANO) generating system using rat bladder. Results suggest that a small number of NADPH-diaphorase-positive perikarya are present within the bladder wall and within adjacent small ganglia. Furthermore, NADPH-diaphorase-positive nerve fibers were observed in the adventitial and muscular layers, subjacent to the urothelium and perivascular fibers. Rat bladder strips precontracted with 3$\mu$M carbachol were reversibly relaxed upon NO generation by UV irradiation. PIANO-mediated relaxation was sensitive to oxygen free radicals. In addition, tissue cGMP levels were increased by the PIANO generating system and elevated cGMP levels were decreased by pretreatment of guanylate cyclase inhibitor, methylene blue. These results indicate that NO may serve a role in modulating bladder tone in the rat.

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Pharmacological Actions of $\imath$--Muscone on Cardiovascular System ($\imath$--Muscone의 실험관계에 관한 약리연구)

  • 조태순;김낙두;허인회;권광일;박석기;심상호;신대희;박대규
    • Biomolecules & Therapeutics
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    • v.5 no.3
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    • pp.299-305
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    • 1997
  • In order to investigate the pharmacological properties of ι-muscone, effects of ι-muscone and musk were studied on the cardiovascular system with various experimental models. In isolated rat aorta, ι-muscone and musk made the relaxation of blood vessels in maximum contractile response to phenylephrine (10$^{-6}$ M) in endothelium-containing rings of the rat aorta, but not in endothelium-denuded rings. However, ι-muscone and musk in the presence of the inhibitor of NO synthase and guanylate cyclase did not make the relaxation of blood vessels. In spontaneously hypertensive rats (SHRs), ι-muscone and musk slightly reduced blood pressure but significantly decreased heart rate. In the isolated perfused rat hearts, ι-muscone and musk did not affect significantly on LVDP, contractile force, coronary flow and (-dp/dt)/(+dp/dt). These results suggest that ι-muscone and musk have weak cardiovascular effects with relaxation of blood vessel and decrease of heart rate, but without significant cardiac functions.

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Purification and Characterization of the $\beta$-Adrenergic Receptor from Rat Cerebral Cortex

  • 신찬영;김희진;노민수;함경수;강현삼;고광호
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1994.04a
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    • pp.292-292
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    • 1994
  • Catecholamines acting through ${\beta}$-adrenergic receptors regulate a wide range of metabolic activities in mammalian tissue. Of the various receptors coupled to adenylate cyclase, the ${\beta}$-adrenergic receptors are the most extensively characterized and have been purified from both nonmammal ian and mammal inn sources. However, most studies of the molecular properties of ${\beta}$-adrenergic receptors have been confined to peripheral tissues. Less progress has been achieved in characterizing the brain ${\beta}$-adrenergic receptor The goal of the present study was, therefore, to purify and characterize the neurotransmitter receptor proteins. To achieve this goal, the following stepwise experiments were performed. At first, the membrane-bound ${\beta}$-adrenergic receptors were. solubi1ized from brain tissue. Secondly, conditions for affinity chromatography were determined to purify the solubilized receptors effectively. Finally, the large-scale purification was performed and the characteristics of the purified ${\beta}$-adrenergic receptor were examined.

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돼지 난자의 체외수정 시 Adenosine Di-phosphate(ADP) 첨가가 배발달에 미치는 영향

  • 이향흔;박진기;김광식;이창현;이연근;김동훈;장원경;송해범
    • Proceedings of the KSAR Conference
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    • 2002.06a
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    • pp.73-73
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    • 2002
  • 돼지 난포란의 체외성숙, 수정 및 배양에 관한 연구는 생명공학 기술을 도입하기 위한 기반 기술로 최근까지 계속 진행되고 있으나, 돼지 난포란을 이용한 효율적인 수정란 생산은 불규칙적인 웅성전핵 형성율과 높은 다정자 침입율 그리고 체내에서 발달된 배반포에 비하여 적은 세포수는 돼지 체외수정 체계에 있어서 끊임없는 문제점으로 제기되고 있다. 따라서 본 연구에서는 난포란을 이용한 보다 효율적인 수정란 생산을 위하여 돼지난자의 체외수정시 정자의 adenylate cyclase 활성을 조절하여 수정능획득 및 자발적인 첨체반응을 조절하여 정상적인 수정을 돕는 것으로 알려진 ADP를 체외수정 배양액에 첨가했을 때 수정율, 다정자 침입율, 배발달율 및 배반포의 세포수에 미치는 영향을 조사하였다. (중략)

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Analysis of a Sphingosine 1-phosphate Receptor $hS1P_3$ in Rat Hepatoma Cells

  • Im, Dong-Soon
    • The Korean Journal of Physiology and Pharmacology
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    • v.6 no.3
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    • pp.139-142
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    • 2002
  • To examine intracellular signaling of human $S1P_3\;(hS1P_3),$ a sphingosine 1-phosphate (S1P) receptor in plasma membrane, $hS1P_3$ DNA was transfected into RH7777 rat hepatoma cell line, and the inhibition of forskolin-induced cAMP accumulation and activation of MAP kinases by S1P were tested. In $hS1P_3$ transformants, S1P inhibited forskolin-induced activation of adenylyl cyclase activity by about 80% and activated MAP kinases in dose-dependent and pertussis-toxin (PTX) sensitive manners. In oocytes expressing $hS1P_3$ receptor, S1P evoked $Cl^-$ conductance. These data suggested that PTX-sensitive G proteins are involved in $hS1P_3-mediated$ signaling, especially the positive action of S1P in cell proliferation. The potential advantages of rat hepatoma cells for the research of sphingosine 1-phosphate receptor are discussed.

The Alpha Subunit of Go Interacts with Promyelocytic Leukemia Zinc Finger Protein

  • Ghil Sung-Ho
    • Biomedical Science Letters
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    • v.10 no.4
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    • pp.407-413
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    • 2004
  • Heterotrimeric GTP binding proteins (G proteins) transduce signals of a variety of hormones and neurotransmitters. Go is one of the most abundant G proteins in the brain and classified as the Gi/Go family due to their sequence homology to Gi proteins. While the Gi proteins inhibit adenylyl cyclase and decrease the intracellular cAMP concentration, the functions of Go is not clearly understood despite their sequence homology to Gi. The promeylocytic leukemia zinc finger protein (PLZF) is a DNA binding transcription factor and is expressed highly in central nervous system (CNS). Several studies reported that PLZF may be involved in regulation segmentation/differentiation during CNS development. Here, I report that the alpha subunit of Go (Go ) interacts with PLZF. The interaction between Goa and PLZF was verified by using GST pulldown assay and co-immunoprecipitation. Our findings indicate that Goa could modulate gene expression via interaction with PLZF during neuronal or brain development.

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THE NEW FINDING OF A LIGHT DEPENDENT $Ca^{2+}$ CHANNEL AND $Na^+-Ca^{2+}$ EXCHANGER IN THE VERTEBRATE RETINA (II)

  • Kim, Yun-Sook;Jung, Hyuk;Park, Chang-Suck;Woo, Suk-Hyang;Kim, Hyun-Jung;Kim, You-Young
    • Journal of Photoscience
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    • v.3 no.3
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    • pp.133-136
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    • 1996
  • Calcium modulates the activity of guanylate cyclase and plays a key role in dark and light adaptation in the visual system. We have measured the Ca$^{2+}$, K$^+$ and Na$^+$ concentration in dark and light adapted bullfrog's (Rana catesbeiana) vitreous humor by using the atomic absorption spectrophotometer. The calcium concentration of the light adapted bullfrog's vitreous humor was higher than that of the dark adapted bullfrog's vitreous humor. This means that ion activity between the photoreceptor and vitreous humor side is light dependent and we have found that a Ca$^{2+}$ channel and Na$^+$ - Ca$^{2+}$ exchanger exist in the vitreous humor.

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CALCIUM EFFECTS OF VISUAL ADAPTATION IN A VERTEBRATE RETINA (I)

  • ung, Hyuk J
    • Journal of Photoscience
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    • v.3 no.3
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    • pp.127-132
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    • 1996
  • Calcium has a variety of functions in neuron and muscle cells and blood clotting, especially in the visual system where dark adapted rods cotransport with Na$^+$ into the cell. An influx of Ca$^{2+}$ flows out of the cell through the Na$^+$ - Ca$^{2+}$ exchanger. By using a modified Ussing chamber in order to bring in vivo environment close, we have concluded that Ca$^{2+}$ blocks the activity of guanylate cyclase; in consequence, having an effect on the amplitude of electroretinogram (ERG). We suggest that Ca$^{2+}$ moves between the photoreceptor and the vitreous humor by way of certain Ca$^{2+}$ transport mechanisms. Also, the effect of Zn$^{2+}$ in Ca$^{2+}$ - free ringer solution caused an elevation of amplitude in ERG and a reduction of threshold.

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In Silico Identification of 6-Phosphogluconolactonase Genes that are Frequently Missing from Completely Sequenced Bacterial Genomes

  • Jeong, Hae-Young;F. Kim, Ji-Hyun;Park, Hong-Seog
    • Genomics & Informatics
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    • v.4 no.4
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    • pp.182-187
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    • 2006
  • 6-Phosphogluconolactonase (6PGL) is one of the key enzymes in the ubiquitous pathways of central carbon metabolism, but bacterial 6PGL had been long known as a missing enzyme even after complete bacterial genome sequence information became available. Although recent experimental characterization suggests that there are two types of 6PGLs (DevB and YbhE), their phylogenetic distribution is severely biased. Here we present that proteins in COG group previously described as 3-oarboxymuconate cyclase (COG2706) are actually the YbhE-type 6PGLs, which are widely distributed in Proteobacteria and Fimicutes. This case exemplifies how erroneous functional description of a member in the reference database commonly used in transitive genome annotation cause systematic problem in the prediction of genes even with universal cellular functions.