장수풍뎅이(Allomyrina dichotoma) 렉틴의 면역기능 증강효과

전경희,정미연,최수정,이종욱,박원학,조세훈,이승호,정시련,Jeune, Kyung-Hee,Jung, Mi-Yeun,Choi, Soo-Jeong,Lee, Jong-Wook,Park, Won-Hark,Cho, Se-Hoon,Lee, Seung-Ho,Chung, See-Ryun 한국생약학회 2001 생약학회지 Vol.32 No.1

A lectin was purified from Allomyrina dichotoma (ADL) by physiological saline extraction, ammonium sulfate fractionation, anion exchange column chromatography on DEAE Sephadex A-50 and gel filtration column chromatography on Sephadex G-200. Several biochemical properties of ADL were characterized as follows: ADL from gel filtration column chromatography showed single band on SDS-PAGE. ADL agglutinated the erythrocytes of rabbit and human A, B, O, AB. Agglutinability was relatively stable at basic pH, and was stable at temperature below $40^{\circ}C$. Agglutinability was not affected by metal ions and EDTA. This lectin was proved to be a glycoprotein which contains 0.47% of sugars. The molecular weight of ADL was estimated to be 97,000 dalton by SDS-PAGE. By amino acid analysis, ADL exhibited high amounts of aspartic acid. The lectin's immunomodulating effect was measured as cytokine production. The productions of 5 cytokines $(IL-1{\alpha},\;IL-2,\;IL-6,\;IFN{\gamma}\;and\;TNF{\alpha})$ from peripheral blood mononuclear cells were measured by ELISA. The lectin induced the highest secretion of IL-2 at 8 hr, $TNF{\alpha}$ at 4 hr, and $IFN{\gamma}$ at 24hr, respectively. These results suggest that ADL can elicit the production of detectable cytokines from PBMC.

콩과 식물에서 분리정제한 렉틴의 생물물리 화학적 연구(E-PHA 렉틴)

전경희,채영흠,서영아,정시련,JeuneChung, Kyung-Hee,Chae, Young-Heum,Suh, Young-Ah,Chung, See-Ryun 생화학분자생물학회 1983 한국생화학회지 Vol.16 No.1

흰강남콩 (Phaseolus vulgaris)의 추출물을 DEAE Sephadex A-50과 hydroxyapatite column을 통과시켜 적혈구 및 립프구 응집효과를 갖는 EL-PHA를 얻었다. EL-PHA로부터 적혈구 응집효과만 가진 E-PHA을 분리하기 위해 EL-PHA를 4M urea로 처리한 다음 CM-Sephadex column을 통과시켜 림프구 응집력만 가진 L-PHA, 적혈구 응집력만 가진 EI-PHA, 적혈구 응집력과 약간의 림프구 응집력을 가진 ElI-PHA를 얻었다. EI-, EII-PHA의 생물물리화학적 성상을 구명코저 activity test, 전기영동, SDS-전기영동, 당류시험, 면역화학적 실험 등을 수행하였다. 전기영동에서 EL-PHA는 3개의 band를 나타내었고, EI-, EII-PHA는 1개의 뚜렷한 band를 나타내었으며, 분자량은 각각 130,000과 125,000 daltons으로 추정되었다. SDS-PAGE 결과 EI-PHA는 분자량이 약 34,000인 subunits 1종으로 나타났고, EII-PHA는 41,000과 30,000정도의 2종의 서로 다른 subunits로 나타났다. 당류에 의한 activity의 저해 효과는 0.3% D-mannose와 N-acetyl-D-galactosamine의 경우에 현저 하였고, D-galactose, L-fucose, N-acetylneuraminic acid, N-acetyl-D-glucosamine의 경우는 저해 효과가 없었다. 한편, EE-, EII-PHA 당류를 미량 함유한 당단백질임을 알 수 있었다. 면역 화학적인 연구의 하나로 immunodiffusion을 한 결과 EI-PHA는 antigen-antibody 반응을 잘 나타내 주었다. Salt fractionated extracts of Phaseolus vulgaris was purified successively through ion exchange chromatography on DEAE Sephadex A-50, hydroxyapatite column and EL-PHA was obtained. EL-PHA was treated with 4M urea just prior to the application into CM-Sephadex C-50 column. This column was eluted with 10 mM (pH 5.7), 20 mM (pH 6.6) and 30mM (pH 8.0) phosphate buffer, each containing 4M urea. In order to identify isolation of E-and L-PHA, erythroagglutination test with human blood A group and lymphoagglutinating test with mouse spleen lymphocytes were measured on each fraction and found three active fractions, L-PHA, having lymphoagglutinating activity only, EI-PHA, having high erythroagglutinating activity only and EII-PHA, having erythroagglutinating activity along with weak lymphoagglutinating activity. In order to characterize biophysicochemical properties of the EI-and EII-PHA, polyacrylamide disc gel electrophoresis, SDS gel electrophoresis, sugar inhibition test, carbohydrate test were performed. In polyacrylamide disc gel electrophoresis, EL-PHA was observed as three bands, EI-and EII-PHA were observed as clear single band with a very faint minor band. Molecular weight of EI-and EII-PHA were estimated at about 130,000 daltons and 125,000 daltons, respectively by PAGE. The results of SDS-PAGE shown that EI-PHA consists of four homogenous subunits having molecular weight of 34,000 per unit and EII-PHA consists of two different subunits with 41,000 arid 30,000 daltons. Erythroagglutination by EI-and EII-PHA was inhibited with 0.3% D-mannose and N-acetyl-D-galactosamine, but not with N-galactose, L-fucose, N-acetylneuraminic acid and N-acetyl-D-glucosamine. Sugar components of EI-and EII-PHA were recognized by anthrone test, but no spot was observed in paper chromatography. For immunochemical studies, antisera to EI-PHA were prepared by immunizing rabbit and EI-PHA formed precipitin bands against various antisera in Ouchterlony double immunodiffusion.

해양동물 렉틴 ( 제 11 보 ) : 조각매물고둥 렉틴의 림프구 분열효과 및 면역화학적 특성

전경희,서영아,소명숙,정시련 ( Kyung Hee Jeune,Young Ah Suh,Myung Suk So,See Ryun Chung ) 생화학분자생물학회 1991 BMB Reports Vol.24 No.3

Developing new substances for immunosuppressor or immunomodulator from natural products is extremely important in the present biomedicine. In this paper, we focused our efforts on the mitogenicity and immunochemical properties of the NI lectins (NIA, NIB, NIC), obtained from Neptunea intersculpta. Immunochemical techniques were employed to elucidate the structural and/or functional similarities between the NI lectins. According to these results, NIB and NIC were almost identical and the NIC proved to be homogeneous on immunoelectrophoresis as well as crossed immunoelectrophoresis. But these lectins are turned out as different molecules through the study of carbohydrate specificity. Only NIC lectin was a potent mitogenic toward murine splenic lymphocytes at a concentration ca. 6.4 ㎍/㎖.

콩과 식물에서 분리정제한 렉틴의 생물물리 화학적 연구 ( E - PHA 렉틴 )

전경희,채영흠,서영아,정시련 ( Kyung Hee Jeune - Chung,Young Heum Chae,Young Ah Suh,See Ryun Chung ) 생화학분자생물학회 1983 BMB Reports Vol.16 No.1

Salt fractionated extracts of Phaseolus vulgaris was purified successively through ion exchange chromatography on DEAE Sephadex A-50, hydroxyapatite column and EL-PHA was obtained. EL-PHA was treated with 4M urea just prior to the application into CM-Sephadex C-50 column. This column was eluted with 10 mM (pH 5.7), 20 mM (pH 6.6) and 30mM (pH 8.0) phosphate buffer, each containing 4M urea. In order to identify isolation of E-and L-PHA, erythroagglutination test with human blood A group and lymphoagglutinating test with mouse spleen lymphocytes were measured on each fraction and found three active fractions, L-PHA, having lymphoagglutinating activity only, EI-PHA, having high erythroagglutinating activity only and EII-PHA, having erythroagglutinating activity along with weak lymphoagglutinating activity. In order to characterize biophysicochemical properties of the EI-and EII-PHA, polyacrylamide disc gel electrophoresis, SDS gel electrophoresis, sugar inhibition test, carbohydrate test were performed. In polyacrylamide disc gel electrophoresis, EL-PHA was observed as three bands, EI-and EII-PHA were observed as clear single band with a very faint minor band. Molecular weight of EI-and EII-PHA were estimated at about 130,000 daltons and 125, 000 daltons, respectively by PAGE. The results of SDS-PAGE shown that EI-PHA consists of four homogenous subunits having molecular weight of 34, 000 per unit and EII-PHA consists of two different subunits with 41,000 arid 30,000 daltons. Erythroagglutination by EI-and EII-PHA was inhibited with 0.3% D-mannose and N-acetyl-D-galactosamine, but not with N-galactose, L-fucose, N-acetylneuraminic acid and N-acetyl-D-glucosamine. Sugar components of EI-and EII-PHA were recognized by anthrone test, but no spot was observed in paper chromatography. For immunochemical studies, antisera to EI-PHA were prepared by immunizing rabbit and EI-PHA formed precipitin bands against various antisera in Ouchterlony double immunodiffusion.

사람 말초혈액 단핵세포에서 녹두 렉틴의 사이토카인 생성효과

전경희,안몽기,정수민,최경민,이승호,정시련,Jeune, Kyung-Hee,An, Mong-Gi,Jung, Su-Min,Choi, Kyung-Min,Lee, Seung-Ho,Chung, See-Ryun 한국생약학회 1999 생약학회지 Vol.30 No.4

New lectins have been isolated and purified from mung bean (Phaseolus radiatus) through physiological saline extraction, ammonium sulfate salt fractionation and column chromatographies. Ion exchanger were eluted by linear salt gradient and then further purified through gel filtration. Thus obtained lectin named as MBL. The gene expressions of 5 cytokines (IL-1, IL-2, IL-6, $TNF-{\aphpa}$ and $IFN-{\gamma}$) from human peripheral blood mononuclear cells (PBMC) stimulated with MBL were investigated by using reverse transcription polymerase chain reaction (RT-PCR). PBMC ($1{\times}106$ cells/ml) isolated from healthy volunteers were stimulated with lectins (4 mg/ml) for various time intervals (1 to 96 hrs). After each of the various stimulated times, total RNA was isolated and assessed for different cytokines mRNA by RT-PCR. The mRNA encoding IL-1, IL-2 were detected continuously from 1 to 20 hrs, and IL-6 was detected up to 24 hrs. But the mRNA encoding $IFN-{\gamma}$ and $TNF-{\alpha}$ were detected to 8 hours only and showed short time response compared with other cytokines. The significant expression of all cytokines mRNA were observed at 4 hrs. These results suggested that MBL, as inducer of cytokines could elicit detectable cytokine mRNA from PBMC.

두유 식물 렉틴의 면역학적 연구

정시련(See Ryun Chung),전경희(Kyung Hee Jeune-Chung) 한국생약학회 1984 생약학회지 Vol.15 No.1

嶺南大學校 藥草園에서 수집한 15種의 콩과植 物種子를 대상으로 사람의 血液과 토끼 血液으로 檢索試驗한 結果 7種 以上의 各種 血液에 대해서 렉틴(lectin)活性을 나타내었다. 이중 5種을 선정하여 電氣泳動試驗結果 各各은 많은 band를 나타내어 이들은 많은 종류의 蛋白質로 되었음을 알 수 있었으며 蛋白質定量結果 多樣한 차이가 나타났다. 한편 이들 콩과植物 렉틴사이에 서로 構造物 相關性이 있는지를 알아보기 위해 免疫生化學的인 硏究를 遂行했든 바, 몇종의 경우는 PHA, E-PHA, L-PHA, MBLA 렉틴등과 免疫生化學的으로 類似한 것으로 밝혀졌으며 더욱 ion exchange chromatography로 分離精製한 PHA에는 면역학적인 실형결과 2종의 抗原이 存在함을 알 수 있었다. 以上의 實驗에서 演者등은 數種 콩과植物을 대상으로 이들의 렉틴含有與否와 PHA, MBLA 렉틴을 이용하여 얻은 antisera로 免疫化學的 實驗을 하여 PHA, MBLA렉틴과 기타 콩과 植物렉틴과의 蛋白質構造내지 機能上의 相關性등을 연구 검토하였다.

두가식물로부터 새로운 렉틴의 분리 및 정제

정시련(See-Ryun Chung),전경희(Kyung-Hee Jeune Chung),김경애(Kyong-Ae Kim) 한국생약학회 1981 생약학회지 Vol.12 No.1

White kidney bean (Phaseolus vulgaris, C, W. K.B.) and Cercis chinensis(C.C) belonging to the Leguminosae contained strong hemagglutinating proteins. The crude lectin from W.K.B. were purified by DEAE Sephadex A-50 column chromatography. The 0.1M peak from this procedure demonstrated acceptable criteria, e.g., yield, optical density and hemagglutinating activity and three bands were observed in polyacrylamide disc gel electrophoresis. Further purification was achieved by hydroxyapatite column chromatography and strong hemagglutinating activity were recovered in 0.1 M peak. I was electrophoresed and found to migrate as a single band. These suggests that the condition of purification for W.K.B. lectin was encourageable but these methods did not brought agreeable satisfaction to the Cercis chinensis lectin.

식물의 렉틴 성분 스크리닝

정시련(See Ryun Chung),정수민(Su Min Chung),이승호(Seung Ho Lee),전경희(Kyung Hee Jeune) 대한약학회 1996 약학회지 Vol.40 No.4

The erythrocytes agglutination test was applied to the common korean plants for lectin activity screening by using human blood. During the four years, 108 species from 46 families of floras were collected, identified and subjected to the test after being divided into several different parts. Only 13 species demonstrated strong lectin activities. Meanwhile 66 species did not shown any agglutination. All others were observed as having low activity or as having hemolytic constituents.

해양패류로부터 렉틴성분 개발연구 2 : 번각매물고동 렉틴의 분리 , 정제 및 특성

정시련(See Ryun Chung),김장환(Jang Hwan Kim),전경희(Kyung Hee Jeune Chung) 영남대학교 자원문제연구소 1987 資源問題硏究 Vol.6 No.-

해양 패류 렉틴성분 연구(V) 반지락조개의 렉틴성분 분리정제에 관한 연구

정시련(See Ryun Chung),김장환(Jang Hwan Kim),전경희(Kyung Hee Jeune-Chung) 대한약학회 1987 약학회지 Vol.31 No.2

The result of the screening of lectins on 28 species of marine shell fishes (mollusks) showed that 10 species (Tapes philippinarum, Cyclosunetta menstrualis, Neptunea arthritica cumingii, Omphalius pfeifferi carpenteri, Chlorostoma argyrostoma turbinatum, Chiorostoma argyrostoma lischkei, Semisulcosira corea, Neptunea polycosta, Babylonica japonica, Noverita didyma) were present hemagglutinating properties to human A, B, and 0 group, and animal blood erythrocytes. A new lectin from Tapes philippinarum was isolated and partially characterized. The lectin was purified by (NH4)2SO4 precipitation and ion exchange chromatography on DE 53 column. Six fractions were obtained from DE 53 column by salt gradient elution but only 0.3M, and 0.4M NaCl fraction had strong lectin activity. On its 0.3M NaCl fraction, purity was identified by polyacrylamide gel electrophoresis. This lectin was inhibited by N-acetyl-D-galactosamine. It seems that two kinds of lectin react as antigen by immunochemical studies.