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Lee, Byoung Dae,Kim, Soomi,Hur, Eun-Mi,Park, Yong-Soo,Kim, Yun-Hee,Lee, Taehoon G.,Kim, Kyong-Tai,Suh, Pann-Ghill,Ryu, Sung Ho Blackwell Science Ltd 2005 Journal of Neurochemistry Vol.95 No.1
<P>Abstract</P><P>Endogenous opioid peptides, found in the central and peripheral nervous systems, perform neuromodulatory roles, and display a wide range of functional and pharmacological properties <I>in vitro</I> and <I>in vivo</I>. In this study, we investigated the effects of prodynorphin gene products on intracellular signaling events and cell survival in rat pheochromocytoma PC12 cells. Leumorphin, but not other prodynorphin gene products including dynorphin A, &bgr;-neoendorphin and rimorphin (dynorphin B), increased cell viability in PC12 cells. The cytoprotective effect of leumorphin was dependent on the phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways, but was insensitive to both naloxone, a general antagonist of the opioid receptor, and nor-binaltorphimine, a specific antagonist of the kappa opioid receptor. Moreover, a competition-binding assay clearly revealed that leumorphin had another binding site(s) in addition to that for the kappa opioid receptor. Interestingly, leumorphin induced activation of the epidermal growth factor receptor via a Src-dependent mechanism, which was proved to be responsible for the increased survival response. Flow cytometric and microscopic analysis showed that leumorphin rescued cells from serum deprivation-induced apoptosis. Collectively, we suggest that leumorphin prevents apoptosis via epidermal growth factor receptor-mediated activation of the phosphatidylinositol 3-kinase and mitogen-activated protein kinase pathways, which occur independent of the kappa opioid receptor.</P>
Ligand profiling and identification technology for searching bioactive ligands
Baek, Moon-Chang,Kim, Sun-Jin,Yea, Kyungmoo,Kim, Youndong,Lee, Byung-Dae,Kim, Jaeyoon,Lee, Hae-Jeong,Kang, Mee-Hee,Choi, Sun-Kyu,Kim, Jong-In,Lee, Taehoon G.,Suh, Pann-Ghill,Ryu, Sung Ho WILEY-VCH Verlag 2006 Proteomics Vol.6 No.6
<P>We introduce a new methodology named ligand profiling and identification for effective discovery of bioactive ligands such as peptide hormones. This technology was developed from a new concept of parallel column chromatography and active fraction profiling by nano-LC MS. Traditional methods use sequential column chromatography, and thus are inevitably limited by the low abundance of the peptide of interest and by a low yield due to the many column steps. Using this new technology, insulin was successfully identified and diarginylinsulin, a minor intermediate form of insulin, was unexpectedly also identified simultaneously from 100 mg of porcine pancreatic tissue. This integrative technology could be used to search for various low-abundance peptides (or bioactive molecules) rapidly and simultaneously, by applying this to the later stages of traditional sequential purification.</P>
Kim, Sun-Hee,Henry, Ellen C,Kim, Dong-Kyu,Kim, Yun-Hee,Shin, Kum Joo,Han, Myoung Sook,Lee, Taehoon G,Kang, Jong-Ku,Gasiewicz, Thomas A,Ryu, Sung Ho,Suh, Pann-Ghill American Society for Pharmacology and Experimental 2006 Molecular pharmacology Vol.69 No.6
<P>2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a widespread environmental pollutant with many toxic effects, including endocrine disruption, reproductive dysfunction, immunotoxicity, liver damage, and cancer. These are mediated by TCDD binding to and activating the aryl hydrocarbon receptor (AhR), a basic helix-loop-helix transcription factor. In this regard, targeting the AhR using novel small molecule inhibitors is an attractive strategy for the development of potential preventive agents. In this study, by screening a chemical library composed of approximately 10,000 compounds, we identified a novel compound, 2-methyl-2H-pyrazole-3-carboxylic acid (2-methyl-4-o-tolylazo-phenyl)-amide (CH-223191), that potently inhibits TCDD-induced AhR-dependent transcription. In addition, CH-223191 blocked the binding of TCDD to AhR and inhibited TCDD-mediated nuclear translocation and DNA binding of AhR. These inhibitory effects of CH-223191 prevented the expression of cytochrome P450 enzymes, target genes of the AhR. Unlike many known antagonists of AhR, CH-223191 did not have detectable AhR agonist-like activity or estrogenic potency, suggesting that CH-223191 is a specific antagonist of AhR. It is noteworthy that CH-223191 potently prevented TCDD-elicited cytochrome P450 induction, liver toxicity, and wasting syndrome in mice. Taken together, these results demonstrate that this novel compound, CH-223191, may be a useful agent for the study of AhR-mediated signal transduction and the prevention of TCDD-associated pathology.</P>
마황으로부터 췌장 Cholesterol Esterase 저해물질 분리 및 규명
조은정(Eun-Jung Cho),류병호(Byung-Ho Ryu),송병권(Byung-Kwon Song),이태훈(Taehoon G. Lee),서판길(Pann-Ghill Suh),류성호(Sung-Ho Ryu),김희숙(Hee-Sook Kim) 한국식품영양과학회 1999 한국식품영양과학회지 Vol.28 No.4
췌장으로부터 분비되는 cholesterol esterase(pCEH, pancreas cholesterol ester hydrolase, E.C.3.1.1.13)는 콜레스테롤 흡수에 관련되는 중요한 지질가수분해효소로 알려져 있다. 식이로부터 섭취되는 cholesteryl acyl ester(이하 콜레스테롤 에스테르)는 소장에서 흡수되기 전에 cholesterol esterase에 의하여 유리콜레스테롤과 지방산으로 가수분해되어야 한다. 천연물질들로부터 동맥경화 또는 고지혈증의 예방제 및 치료제를 개발할 목적으로 본 실험실에서는 여러 가지 한약재료의 추출물들을 이용하여 cholesterol esterase에 대한 저해제활성을 시험관법으로 검색하였는데 마황의 에탄올추출물이 강한 저해활성을 보였다. 마황의 에탄올추출물로부터 용매분획법과 silica gel column chromatography 등을 이용하여 cholesterol esterase활성을 저해하는 물질을 분리 정제하였으며 저해활성을 가진 흰색 결정은 UV, ¹H NMR, ^(13)C-NMR 및 GC/Mass로 분석한 결과(-)-ephedrine으로 동정되었다. 이러한 결과들은 (-)-ephedrine이 콜레스테롤 에스터레이즈를 저해하므로서 식이중 콜레스테롤의 흡수를 저해하는 흡수저해제를 개발하기 위한 기본 물질로 이용될 수 있을 것으로 생각된다. Cholesterol esterase(pCEH, pancreas cholesterol ester hydrolase, E.C.3.1.1.13) which is secreted from pancreas has been known as an important lipase for cholesterol uptake. cholesteryl acyl esters from a diet must be hydrolyzed to free cholesterol and fatty acid by cholesterol esterase before the absorption in small intestine. For the development of inhibitory substances from natural source, we screened many extracts of oriental herbs for the inhibition of cholesterol esterase in vitro. The ethanol extract of Ephedra herba showed strong inhibitory activity. Solvent fractionation and silica gel column chromatography with the extract lead to the purification of the inhibitory principle in Ephedra herba. Crystallized inhibitor was identified as (-)-ephedrine by using UV, FT-IR, ¹H-NMR, ^(13)C-NMR and GC/Mass. These results suggest that (-)-ephedrine can be used as a potential lead compound for the development of inhibitor for cholesterol uptake by cholesterol esterase inhibition.