Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology ( SH ) 2 Domain and Phosphotyrosine Interactions

(Sang Seop Lee),(Kyung Im Lee),(Ji Yun Yoo),(Moon Jin Jeong),(Young Mee Park),(Byoung Mog Kwon),(Yun Soo Bae),(Mi Young Han) 생화학분자생물학회 2001 BMB Reports Vol.34 No.6

In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/ SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/ SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.

Isolation and Characterization of Marinobacter sp. KS-1, which Produces a Chondroitin Sulfate-like Mucopolysaccharide

Lee, Dae-Sung,Kim, Kyung-Suk,Lee, Myung-Suk,Kim, Young-Mog The Korean Society of Fisheries and Aquatic Scienc 2010 Fisheries and Aquatic Sciences Vol.13 No.3

In an effort to isolate a bacterium producing chondroitin sulfate (CS), a marine bacterium, KS-1, which produces mucopolysaccharides, was isolated from seawater and identified as Marinobacter sp. based on analyses of its morphological and biochemical traits and 16S rDNA sequence. Agarose-gel electrophoresis showed that the KS-1 strain produces a CS-like mucopolysaccharide. Structural analysis using Fourier transform infrared spectroscopy revealed that the structure of the CS-like mucopolysaccharide produced by Marinobacter sp. KS-1 is similar to that of dermatan sulfate (CS B). However, the molecular mass of the CS-like mucopolysaccharide is higher than that of standard chondroitin sulfates. Considering the above results, we conclude that the Marinobacter sp. KS-1 produces a CS-like mucopolysaccharide that differs somewhat from CS B in molecular mass.

The Representation of the Neolithic Rock Art of Bangu-dae, South Korea

Lee, Sang-Mog Korea Association For Quaternary Research 2004 제사기학회지 Vol.18 No.2

The rock engraving of the Korea are dated by a very few direct archaeological contexts by linking their images to objects of known prehistoric date-although many of the figures are 'abstract'. This paper focuses on the rock art of Bangu-dae, located in the south-east of Korea. I try to the date of engravings from their contexts and the ways in which they can be studied.

Short Communications : Article ; Radical Scavenging Activity of Poncirus trifoliata Extracts and Their Inhibitory Effect Against Hydrogen Peroxide Induced Cell Damage

( Sung Hwan Eom ),( Soo Jin Heo ),( Dae Sung Lee ),( Myung Suk Lee ),( Young Man Kim ),( Won Kyo Jung ),( Young Mog Kim ) 한국응용생명화학회(구 한국농화학회) 2011 Applied Biological Chemistry (Appl Biol Chem) Vol.54 No.3

Poncirus trifoliata was evaluated for antioxidant potential by several radical-scavenging assays, including 1,1-diphenyl-2-picrylhydrazyl, superoxide, hydroxyl, and alkyl radicals, using electron spin resonance technique. P. trifoliata extracts were prepared by using water and methanol, partitioning with different organic solvents. Among the tested samples, water extract and dichloromethane fraction evidenced relatively higher scavenging activities than those of other samples on reactive radicals and have high amount of antioxidant component such as flavonoids (8.5±0.3 and 8.3±0.2 mg rutin equivalents/g dry weight basis) and proanthocyanidin (2.6±0.3 and 2.8±0.5 mg catechin equivalents/g dry weight basis), respectively. Moreover, the water extract showed significantly high enhancement of cell viability against hydrogen peroxide-induced oxidative damage as well as reduction in the hydrogen peroxide-induced intracellular reactive oxygen species on CCL 13 cell line. These results indicate that P. trifoliate exerts various antioxidant effects against several reactive radicals and hydrogen peroxide-mediated cell damage. Thus, P. trifoliate could be a potential therapeutic modality for the treatment or prevention of several diseases associated with oxidative stress.

Synergistic Antibacterial Effect and Antibacterial Action Mode of Chitosan-Ferulic Acid Conjugate against Methicillin-Resistant Staphylococcus aureus

( Sung-hwan Eom ),( Shin-kook Kang ),( Dae-sung Lee ),( Jeong-in Myeong ),( Jinhwan Lee ),( Hyun-woo Kim ),( Kyoung-ho Kim ),( Jae-young Je ),( Won-kyo Jung ),( Young-mog Kim ) 한국미생물 · 생명공학회 2016 Journal of microbiology and biotechnology Vol.26 No.4

We evaluated the synergistic antibacterial effect in combination with the chitosan-ferulic acid conjugate (CFA) and β-lactam antibiotics, such as ampicillin, penicillin, and oxacillin, against methicillin-resistant Staphylococcus aureus (MRSA) using fractional inhibitory concentration (FIC) indices. CFA clearly reversed the antibacterial activity of ampicillin, penicillin, and oxacillin against MRSA in the combination mode. Among these antibiotics, the combination of oxacillin-CFA resulted in a ΣFICmin range of 0.250 and ΣFICmax of 0.563, suggesting that the oxacillin-CFA combination resulted in an antibacterial synergy effect against MRSA. In addition, we determined that CFA inhibited the mRNA expression of gene mecA and the production of PBP2a, which is a key determinant for β-lactam antibiotic resistance, in a dosedependent manner. Thus, the results obtained in this study supported the idea on the antibacterial action mechanism that oxacillin will restore the antibacterial activity against MRSA through the suppression of PBP2a production by CFA.

Symboles et Signes des Sources Arch?ologiques - les Th?mes Iconographiques de Repr?sentation Pr?historique dans la Cor?e : 고고학 유물 속에 나타나는 상징ㆍ기호에 대한 연구

Sang-Mog LEE(이상목) 한국고대학회 2006 先史와 古代 Vol.25 No.-

이 글에서는 주로 우리나라 신석기시대에서 청동기시대 사이의 유적과 유물들 속에 나타난 상징과 기호들을 분석하고, 제작시대와 그 변화양상에 대해 살펴보았다. 연구 대상은 크게 사실적인 형상과 추상적인 형상으로 구분 할 수 있으며, 이는 유적의 성격과 시대적 차이에서 비롯된 것으로 판단할 수 있다. 즉, 수렵채집경제 단계에 해당하는 신석기시대의 조형물에서는 사실적이고도 다양한 동물 표현물이, 농경단계로 진입하는 청동기시대에서는 특정한 주제의 새로운 추상적 표현물들이 나타난다. 이러한 분석을 통해서 상징과 기호 대한 사회경제적인 의미와 당시의 생활상의 변화를 추론하였다. 본 연구를 통하여 사실적인 형상은 신석기시대의 수렵어로 경제단계의 소산물로, 사냥꾼사회의 계절적 식량사원 취득과 생업활동을 잘 반영한 것으로 여겨진다. 반면 청동기시대에 접어들면서 새로운 양식의 형상들이 출현하게 되며, 매우 추상적인 주제(방패, 동심원, 검, 여성음부 등)로 한정되어 있다. 이러한 상징의 변화는 농경이란 새로운 생활방식과 사회구조의 변화에 매우 중요한 역할을 하였을 것으로 추론할 수 있다. In a context archeological, we can gather into two categories theme of representations prehistory: that is to say the representation naturalist is the abstract figuration. The sites that have naturalistic representations probably relate to an early, pre-agricultural way of life. Others seem later - agricultural - as, for example, the site linked to megalithic culture. They can be dated by a very few direct stratigraphic contexts and by linking their images to objects of known prehistoric date, although many of the figures art ‘abstract’ and cannot be dated that way. One can be dated indirectly, a plethora of whale and seal bones and harpoons from a nearby shell midden, that suggests the people who lived there 7,000 to 3,500 years ago. Others are found on megalithic formations, particularly dolmens and menhirs. A close correlation has been shown to exist between the distribution of archeological and symbolic sites in this case. These sites of engravings probably belong to the Bronze Age (between 1,000 and 300BC). According to this analysis, we can suggest certain conclusions on the level of cultural development reached, living conditions, religious observance, etc. We can distinguish at least two different stages; one of the fishing-hunting and the other of agriculture. The majority of the representations of animals by the populations of hunters-fishers are rather close to reality, while the farming populations had an abstracted art symbolic system. In the populations of fishers-hunters, the realism can have a high value symbolic system. On the other hand, the agricultural societies are limited to some geometrical figurations and arms: circles, vulvas, daggers, etc. The suggestion concerning chronology may be further substantiated through their relationship to context. Rock-art has had a function in society and will therefore relate in some form to social structure. In conclusion, one global response to the result of environmental and technological change has been a creative explosion for inspiration on cultures the world over; not only in technology, but also in the arts . In other words, the hunter-fishers’ rock-art tradition came to an end and the symbolic expression of a new economy and ideology took its place. A symbol functioned as an important agent in legitimizing and mediating status and inequality in a period of stressful social transition.

An Experimental Study on the Efficacy of Doxycycline Hydrochloride and Thiamphenicol Actinobacillus Pleuropneumonia Infection in Pigs

Lee, Sang-Mog,Lee, Won-Chang,Sakai, Takeo,Hamakawa, Masaaki,Yoon, Chong-sam,Masuda, Testuya 건국대학교 동물자원연구센터 1991 動物資源硏究誌 Vol.16 No.-

Actinobacillus Pleuropneumonia(AP)에 感染된 豚에게 doxycycline hydrochloride(DOXY)와 thiamphenicol(TP)를 각각 100ppm에서 200ppm씩 食餌에 첨가하여 7일간 投與한 결과에서는 기침(咳)과 鼻漏의 치료효과가 관찰되었으며, 17일 후에서는 體重의 증가와 辭料攝取率이 向上되었고, 部檢上 肺藏에 病變이 없었으며, 겸하여 DOXY의 100ppm과 200ppm 投與群에서는 肺臟에서 Actinobacillus Pleuropneumonia와 KPasteurella multocida가 분리되지 않았다. In order to an experimental find out on the antimicrobial efficacy of doxycycline hydrochloride(DOXY) and thiamphenicol(TP) were given to pigs infected with Actinnobacillus pleuropneumonia(AP) for 7 days. Clinical therapeutic efficacy on coughing and rhinorrhea was observed with 100ppm to 200ppm in the diet. Weight gain and food consumption were improved and pulmonary lesions were not observed at necropsy in the 100 ppm or 200 ppm groups on day 17. In addition, neither Ap nor Pasteurella multocida(Pm) was isolated from the lungs of pigs in the 100 ppm or 200 ppm DOXY on day 17. Key word: antimicrobial efficacy of DOXY and TP.

Study on the Mercury Contents in Korean Scalp Hair : (第2報) 毛髮中 總水銀含量과 메칠水銀量間의 相關 考察 (Report Ⅱ) An Observation on the Rlationship between Total Mercury and Methylmercury Contents in the Scalp Hair

Lee, Won-Chang,Lee, Sang-Mog 건국대학교 1990 學術誌 Vol.34 No.2

저자들은 모발중에 총수은함량과 메칠수은함량간의 상관성을 검토하기 위하여 서울 시민 중 남녀 315명의 모발을 대상으로 하여 정량분석한 격과(제1보)에 이어 제2보로서 본 연구를 시도한 바 다음과 같이 그 결과를 요약할 수 있었다. 연구대상 중 남성의 모발은 여성들의 모발 중에 총수은량과 메칠수은의 함량에 비하여 통계학적으로 유의하게 높았고(p<0.01), 그러나 극히 일부를 제외하고는 안전권 범위에 있어 공중보건학상 이직까지는 문제되지 않고 있었다. 그리고 모발중 총수은함량과 메칠수은량간의 통계학적 상관도를 검사한 바 남성들의 모발에서는 r=+0.862(p<0.01)과 여성 모발에서는 r=+0.721(p<0.01) 그리고 남녀양성을 통합하여 본 바는 r=+0.941(p<0.01)로 각각 높은 상관성이 있어, 이를 이용하여 모발 중의 총수은함량에 대한 예측되는 메칠수은량을 산출할 수 있는 회귀방정식을 각각 고안하여 보았다. The authors conducted an observative study on the relationship between total mercury (TM) and methylmercury (MM) contents in trace metal level of Korean scalp hair samples, which never been made before in the field of Environmental Health and Toxicology in Korea. The following results were summerized: A higher level of total mercury and methylmercury amount has been found in the scalp hair of Korean male than female (p<0.01). The statistical coefficient of correlation between total mercury and methylmercury contents in scalp hair of male was r=+0.862(p<0.01) and that of female was r=+0.721 (p<0.01) and that of sum in male and female was r=+0.941 (p<0.01), respectively.

Heyneanol A induces apoptosis via cytochrome c release and caspase activation in human leukemic U937 cells

Lee, Eun-Ok,Kwon, Byoung-Mog,Song, Gyu-Yong,Chae, Chan-Hee,Kim, Hyung-Min,Shim, In-Sop,Ahn, Kyoo-Seok,Kim, Sung-Hoon WHO COLLABORATING CENTRE FOR TRADITIONAL MEDICINE 2004 東西醫學硏究所 論文集 Vol.2004 No.-

Heyneanol A, a tetramer of resveratrol, is isolated from the roots of Vitis amurensis by cytotoxicity based fractionation. In this study, the mechanism of apoptosis by heyneanol A was evaluated in human leukemic U937 cells. Heyneanol A (IC_(50) = 6.6 μM at 24 h) exhibited stronger cytotoxic effect than resveratrol (IC_(50) = 100 μM at 24 h) by 15-fold an human leukemic U937 cells by XTT assay. Apoptotic bodies were observed in U937 cells treated with 6 μM of heyneanol A by TUNEL assay. Heyneanol A effectively increased the Portion of sub-G₁ DNA content in a time- and concentration-dependent manner by flow cytometric analysis. Heyneanol A also induced cytochrome c release from mitochondria into the cytosol and subsequent caspase activation involving caspase 9 and 3 to cleave PARP. However, it did not affect the expressions of Bax and Bcl-2 by western blotting. It was confirmed that the activation of caspase 8, 9 and 3 and the cleavage of PARP by heyneanol A were completely blocked by adding Z-VAD-FMK, a caspase inhibitor. These findings suggest that heyneanol A has anti-tumor activity, which may be mediated by apoptosis caused by cytochrome c release and caspase activation in human leukemic U937 cells.

Development of ELISA System for Screening of Specific Binding Inhibitors for Src Homology (SH)2 Domain and Phosphotyrosine Interactions

Lee, Sang-Seop,Lee, Kyung-Im,Yoo, Ji-Yun,Jeong, Moon-Jin,Park, Young-Mee,Kwon, Byoung-Mog,Bae, Yun-Soo,Han, Mi-Young Korean Society for Biochemistry and Molecular Biol 2001 Journal of biochemistry and molecular biology Vol.34 No.6

In the present study, an in vitro ELISA system to assess the interaction between Src homology (SH)2 domains and phosphotyrosine that contain peptides was established using purified GST-conjugated SH2 proteins and synthetic biotinylated phosphotyrosine that contain oligopeptides. The SH2 domains bound the relevant phosphopeptides that were immobilized in the streptavidin-coated microtiter plate in a highly specific and dose-dependent manner. The epidermal growth factor receptor (EGFR)-, T antigen (T Ag)-, and platelet-derived growth factor receptor (PDGFR)-derived phosphopeptides interacted with the growth factor receptor binding protein (Grb)2/SH2, Lck/SH2, and phosphatidyl inositol 3-kinase (PI3K) p85/SH2, respectively. No cross-reactions were observed. Competitive inhibition experiments showed that a short phosphopeptide of only four amino acids was long enough to determine the binding specificity. Optimal concentrations of the GST-SH2 fusion protein and phosphopeptide in this new ELISA system for screening the binding blockers were chosen at 2nM and 500nM, respectively. When two candidate compounds were tested in our ELISA system, they specifically inhibited the Lck/SH2 and/or p85/SH2 binding to the relevant phosphopeptides. Our results indicate that this ELISA system could be used as an easy screening method for the discovery of specific binding blockers of protein-protein interactions via SH2 domains.