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Detection and Characterization of Extracellular Phospholipase $A_2$ Activity in Human Amniotic Fluid
백석환,이승호,장현욱,Baek, Suk-Hwan,Lee, Seung-Ho,Chang, Hyeun-Wook 생화학분자생물학회 1992 한국생화학회지 Vol.25 No.3
사람의 양수 중에서 처음으로 세포외성 phospholipase $A_2$ 활성이 있음을 확언하였으며, 출산에 있어서 중요한 prostaglandin의 전구체인 arachidonic acid 제공에 중요한 역활을 할 것이라고 생각된다. 본 효소는 지금까지 보고된 group I이나 group II phospholipase $A_2$와 계변 활성제의 영향이나 monoclonal antibody와의 반응성에서 다른 특성을 갖는 효소로 추정되었으며, 또한 적어도 2종류의 phospholipase $A_2$의 활성이 존재함에 시사되어 금후 더욱 구체적인 검토를 행하여 단백화학적 특성을 규명할 필요가 있다. Extracellular phospholipase $A_2$ activity has been identified in human amniotic fluid. This enzyme required $Ca^{2+}$ ion and exhibited bimodal pH optimum at pH 7.0 and 9.0. The phospholipase hydrolyzed 1-acyl-2-[1-$^{14}C$]arachidonoylphosphatidylethanolamine to form only [$^{14}C$]arachidonic acid indicating that the enzyme had phospholipase "$A_2$" activity. Ionic and non-ionic detergents caused loss of enzymatic activity. This phospholipase $A_2$ was recognized, in part, by a monoclonal antibody raised against phospholipase $A_2$ from human synovial fluid. This finding suggests that our enzyme is an another type of an extracellular phospholipase $A_2$ which may not belong to the 14 KDa group II phospholipase $A_2$ family.
사람 양수 중의 세포외성 Phospholipase A2 특성
백석환,이승호,장현욱 ( Suk Hwan Baek,Seung Ho Lee,Hyeun Wook Chang ) 생화학분자생물학회 1992 BMB Reports Vol.25 No.3
Extracellular phospholipase A₂ activity has been identified in human amniotic fluid. This enzyme required Ca^(2+) ion and exhibited bimodal pH optimum at pH 7.0 and 9.0. The phospholipase hydrolyzed 1-acyl-2-[1-^(14)C]arachidonoylphosphatidylethanolamine to form only [^(14)C]arachidonic acid indicating that the enzyme had phospholipase $quot;A₂$quot; activity. Ionic and non-ionic detergents caused loss of enzymatic activity. This phospholipase A₂ was recognized, in part, by a monoclonal antibody raised against phospholipase A₂ from human synovial fluid. This finding suggests that our enzyme is an another type of an extracellular phospholipase A₂ which may not belong to the 14 KDa group II phospholipase A₂ family. v
연구논문 : 생명과학 ; 금마타리에서 분리한 나르도스타틴의 NO, PGE2 및 TNF 생성 억제효과
주혜경 ( Hye Kyung Ju ),백석환 ( Suk Hwan Baek ),안인파 ( Ren Bo An ),배기환 ( Ki Hwan Bae ),손건호 ( Kun Ho Son ),김현표 ( Hyun Pyo Kim ),강삼식 ( Sam Sik Kang ),이승호 ( Sung Ho Lee ),손종근 ( Jong Keun Son ),장현욱 ( Hyeun Wook 영남대학교 약품개발연구소 2004 영남대학교 약품개발연구소 연구업적집 Vol.14 No.-
사람 양수중 다종의 세포외성 포스포리파제 A2의 부분정제 및 특성
전용주(Yong Ju Jeon),백석환(Suk Hwan Baek),이지혜(Jee Hae Lee),문태철(Tae Chul Moon),민병우(Beong Woo Min),장현욱(Hyeun Wook Chang) 대한약학회 1997 약학회지 Vol.41 No.2
Multiple forms of extracellular phospholipase A2 have been detected in human amniotic fluid (HAF). When HAF was subjected to heparin-Sepharose column chromatography, phospholipase A2 activity was detected in both heparin-non binding and binding fraction. The activity of heparin-non binding fraction was further purified by sequential uses of column chromatographies on butyl-Toy-opearl 650M and DEAE-Sephacel. DEAE-Sephacel fraction contained three different phospholipase A2 activities (Peak I, II, III). The molecular weight of DEAE-Sephacel fraction phospholipase A2 determined by SDS-PAGE were about 52KDa (Peak I). Peak II, III required micromolar Ca2+ ion for its maximum activity, but Peak I enzyme showed calcium independent phospholipase A2 activity and showed broad range of pH (6.0~10.0) optimum. All these enzymes were not recognized by a monoclonal antibody raised against phospholipase A2 from human synovial fluid. These results suggest that HAF might contain multiple forms of extracellular phospholipase A2, which may neither belong to the 14KDa group II phospholipase A2 family nor cytosolic phospholipase A2.
연구논문 : 생명과학 ; 프로페논 화합물의 배양 대식세포에서 NFκ-B활성화 억제를 통한 NO 및 TNF-α 생성억제
이응석 ( Eung Seok Lee ),주혜경 ( Hye Kyung Ju ),문태철 ( Tae Churl Moon ),이은경 ( Eun Kyung Lee ),장영동 ( Yurng Dong Jahng ),이승호 ( Sung Ho Lee ),손종근 ( Jong Keun Son ),백석환 ( Suk Hwan Baek ),장현욱 ( Hyeun Wook Chang 영남대학교 약품개발연구소 2004 영남대학교 약품개발연구소 연구업적집 Vol.14 No.-
Baek, Suk-Hwan,Yun, Sung-Su,Kwon, Taeg Kyu,Kim, Jae-Ryong,Chang, Hyeun-Wook,Kwak, Jong-Young,Kim, Jung-Hye,Kwun, Koing-Bo 영남대학교 약품개발연구소 2000 영남대학교 약품개발연구소 연구업적집 Vol.10 No.-
Phospholipase A_(2)(PLA_(2)) regulates eicosanoid and platelet-activating factor production. It also plays an important role in the regulation of critical mediators in inflammatory diseases in which PLA_(2) activity is significantly enhanced during sepsis and multiple organ failure. Therefore, inhibitors of PLA_(2) activity offer themselves as target substances in the development of anti-inflammatory drugs. We identified 2 biflavonoids, bilobetin and ginkgetin, that can inhibit PLA_(2) activity. In experiments using 2-linol-[1-^(14)C]PE as substrate both substances potently inhibited several kinds of typeⅡ 14-kDa PLA_(2) while inhibiting typeⅠ 14-kDa PLA_(2) to a lesser extent. We tested these PLA_(2) inhibitors for their ability to inhibit the production of tumor necrosis factor α(TNFα) and 2 enzymes, inducible nitric oxide synthase (iNOS) and inducible cyclo-oxygenase(COX-2) in an assay system using lipopolysaccharide(LPS)-stimulated Raw264.7 macrophages. In Raw264.7cells, bacterial LPS induced the production of COX-2 and iNOS proteins as well as TNFα. The inhibitors consistently inhibited the production of TNFα in a dose-dependent manner. Moreover, treatment of the macrophages with bilobetin and ginkgetin shut down the production of nitrite, one of the stable end products of NO released into the culture supernatant. The decrease in NO products was accompanied by a decrease in iNOS protein level as assessed by Western blot probed with specific anti-iNOS antibody. Both inhibitors also reduced the expression of COX-2 protein in the LPS-stimulated cells, which coincided with the reduction in iNOS protein. These results, therefore, suggest that these two sPLA_(2) inhibitors may be useful for inhibiting the production of inflammatory cytokine and NO production in inflammatory diseases.
MOON, Tae Chul,LEE, Jee Hae,LEE, Sung Ho,PARK, Yoon Kee,BAEK, Suk Hwan,CHANG, Hyeun Wook 영남대학교 약품개발연구소 2001 영남대학교 약품개발연구소 연구업적집 Vol.11 No.-
Two types of phospholipase A_(2) (PLA_(2)) Inhibitory proteln (PLIP-Ⅰ, PLIP-Ⅱ) were detected and lsolated from human amniote fluid by Sephacryl S300 gel flitration chromatiography. The lower molecular welght-fractlon (PLIP-Ⅱ) was further purifled by Sephadex G75 ge; fo;tratopm amd ama;uzed bu SDS-PAGE. Its molecular weight was estimated to be approximately 18 kDa. and it was sensitive to heat treatment. Inhibitlon of hposphollpase A_(2) (sPLA_(2) type IIA) by PLIP-Ⅱ oceurred in a dose-dependent manner (IC_(30) about 0.82㎛), and the effect was stronger on sPLA_(2) IIA than on pancreatic sPLA_(2) (IC_(30) about 3.11㎛). The ration of the inhibitions of the sPLA_(2) IIA by PLIP-Ⅱ remalned consistent over an entire range of substrate concentrations. Furthermore, addition of excess Ca^(2+) at comcentrations of up to 10mM did not antagonize the lnhlbitory activity of PLIP-Ⅱ.
정상인의 혈중 및 뇨중 Extracellular Phospholipase A_2 활성도
황윤근,서영익,강문수,최영환,이종명,김보완,김능수,서장수,이주형,배현혜,김경호,이중기,백석환,장현욱 慶北大學校 醫科大學 1993 慶北醫大誌 Vol.34 No.3
Extracellular phospholipase A_2(PLA_2) is known to be secreted by the mononucleocytes, granulocytes, platelet, and chondrocytes in the joint, and it is also reported that this enzyme is increased in inflammatory conditions, such as acute panceratitis, sepsis, ARDS and arthritis. It was proposed that the extracellular release of phospholipase A_2 in response to inflammatory stimuli might represent an amplification mechanism for the generation of high levels of lipid mediators such as eicosanoids. To establish the normal value of PLA_2 in the healthy individuals we measured extracellular PLAz of healthy individuals of 84 males and 85 females in their serum and urine. The levels of PLA_2 activity in serum were 170.85±79.92 (U/㎖) in under 29 years old (20 males), 160.68±109.61 in 30 to 39 years old (19 males), 226.58±130.47 in 40 to 49 years old (19 males), 231.10±82.32 in 50 to 59 years old(17 males), 228.21±102.46 in over 60 years old(6 males), 168.21±62.11(U/㎖) in under 29 years old(6 females), 125.85±101.64 in 30 to 39 years old(20 females), 86.70±31.03 in 40 to 49 years old(20 females), 79.00±35.51 in 50 to 59 years old(20 females), 104.74±46.97 in over 60 years old(19 females). The levels of PLA_2 activity in urine were 142.61±141.86(U/㎖) in under 29 years old(18 males), 377.85±273.01 in 30 to 39 years old(20 males), 406.16±318.07 in 40 to 49 years old(19 males), 84.34±56.12(U/㎖) in under 29 years old(6 females), 17.20±9.94 in 30 to 39 years old(15 females), 54.98±86.24 in 40 to 49 years old(15 females), 14.78±11.38 in 50 to 59 years old(18 females), 43.00±40.29 in over 60 years old(19 females).