• Title/Summary/Keyword: Deoxycholate

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THE AFFINITY OF CALMODULIN-AFFIGEL FOR INOSITOL TRIPHOSPHATE KINASE FROM BOVINE BRAIN (소의 뇌 Inositol triphosphate kinase와 Calmodulin-Affigel과의 친화도)

  • Lim, Sung-Woo;Kim, Jung-Hye
    • Journal of Yeungnam Medical Science
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    • v.7 no.1
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    • pp.39-50
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    • 1990
  • The one event on signalling mechanism is the cleavage by adenyl cyclase of ATP into second messenger, cyclic AMP. The other transfer system of inositol metabolism. it is widely recognized that hydrolysis of the minor membrane lipid phosphoinositide bisphosphate($PIP_2$) initiated by occupation of certain receptors and catalyzed by phospholipase C, lead to toe generation of the two intracellular messengers, inositol triphosphate($IP_3$) and diacylglycerol(DG). $IP_3$ is converted to inositol tetrakisphosphate($IP_4$) by $IP_3$ kinase. In the present study, it is that purification of calmodulin is used by phenyl-Sepharose CL-4B chromatography. it's molecular weigh, 17.000 in SDS-polyacrylamide gel electrophoresis. In order to observe the affinity between calmodulin (CaM)-Affigel 15 and $IP_3$ kinase, and isolated $IP_3$ kinase, was applied in CaM-Affigel with $Ca^{2+}$ equilibirum buffer and EGTA equilibirum buffer. We compared with binding and elution effect of $IP_3$ kinase in several condition of buffer. In affinity of binding. $Ca^{2+}$ equilibrium buffer was in the most proper condition. and elution, CaM/$Ca^{2+}$ buffer(CE1 10.36, CE2 12. 76pM/min/mg of protein) was effected much more than EGTA buffer(E2 1.48, E3 2.43pM/min/mg of protein), but CaM/$Ca^{2+}$ stimulate the activity of $IP_3$ kinase. And then, several detergents such as sodium deoxycholate, tween 20. cholic acid, polyethylene glycol, chaps were applied. The 0.2% chaps buffer(E2 23.19, E3 8.05pM/min/mg of protein) was the most effective in elution of $IP_3$ kinase.

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Rheological Properties of Cement Paste Mixed with Aqueously Dispersed Single-Walled Carbon Nanotubes (Single-Walled 탄소나노튜브 수용액 혼입 시멘트 페이스트의 유변학적 특성)

  • Kim, Ji-Hyun;Chung, Chul-Woo
    • Journal of the Korea Institute of Building Construction
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    • v.19 no.2
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    • pp.113-121
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    • 2019
  • Single walled carbon nanotube (SWCNT) has been used as a material for reinforcing various advanced materials because it has superior mechanical properties. However, pure SWCNT that does not have any functional group has a hydrophobic character, and exists as bundles due to the strong Van der Waals attraction between each SWCNT. Due to these reasons, it is very difficult to disperse SWCNTs in the water. In this work, in order to use SWCNT for production of cementitious composites, SWCNT was first dispersed in water to make an aqueous solution. Sodium deoxycholate (DOC) and Sodium dodecyl sulfate (SDS) were chosen as surfactants, and the dosage of DOC and SDS were 2wt% and 1wt%, respectively. Sonication and ultracentrifugation were applied to separate each SWCNT and impurities. Using such processed SWCNT solutions, cement paste was prepared and its shear stress vs. strain rate relationship was studied. The yield stress and plastic viscosity of cement paste were obtained using Bingham model. According to the results in this work, cement pastes made with DOC and SDS showed similar rheological behavior to that of air entrained cement paste. While cement paste made with DOC 2 wt.% SWCNT solution showed similar rheological behavior to that of plain cement paste, cement paste made with SDS 1 wt.% SWCNT solution showed different rheological behavior showing much less yield stress than plain cement paste.

Biochemical Characterization of Phospholipase C$\delta$from liver of Mud loach (Misgurnus mizolepis) (미꾸라지 간으로부터 포스포리파아제 C델타 단백질의 생화학적 특성)

  • Seo, Jung-Soo;Lim, Sang-Uk;Kim, Na-Young;Lee, Sang-Hwan;Oh, Hyun-Suk;Lee, Hyung-Ho;Chung, Joon-Ki
    • Journal of fish pathology
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    • v.18 no.1
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    • pp.67-80
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    • 2005
  • Phosphoinositide-specific phospholipase $C\delta$ $PLC\delta$) plays an important role in many cellular responses and is involved in the production of second messenger. The present study was conducted to obtain the biochemical characteristics of the expressed recombinant $PLC\delta$ in E. coli cloned from Misgurnus mizolepis and partially purified $PLC\delta$ enzymes from liver tissues of M. mizolepis (wild ML-$PLC\delta$). The ML $PLC\delta$ gene was cloned and expressed under the previous report (Kim et al., 2004), and purified the recombinant protein by successive chromatography using $Ni^{2+}$-NTA affinity column and gel iltration FPLC column. The wild ML-$PLC\delta$ protein was solublized with 2 M KCI and purified by successive chromatography on open heparin-Sephagel and analytical TSKgel heparin-5PW. Both the recombinant and wild ML-$PLC\delta$ form of protein showed a concentration-dependent PLC activity to phosphatidylinositol 4,5-bis-phosphate (PIP$_2$) or phosphatidylinositol (PI). Its activity was absolutely $Ca^{2+}$- dependant, which was similar to mammalian $PLC\delta$ isozymes. Maximal PI-hydrolytic activations of recombinant and wild ML- TEX>$PLC\delta$ was at pH 7.0 and pH 7.5, respectively. In addition, the enzymatic activities of recombinant and wild ML-$PLC\delta$ were increased in concentration-dependent manner by detergent, such as sodium deoxycholate SDC), phosphatidylethanolamine (PE) and phosphatidylcholine (PC). The activities decreased in contrast by a polyamine, such as spermine. Western blotting showed that several types of $PLC\delta$ isozymes exist in various organs. Taken together our results, it suggested that the biochemical characteristics of ML-$PLC\delta$ are similar with those of mammalian $PLC\delta1$ and ${\delta}3$ isozymes.