버섯류의 렉틴성분 개발연구(I): 혈구응집성 단백질의 분리

全瓊姬,鄭時鍊 嶺南大學校附設 基礎科學硏究所 1986 基礎科學硏究 Vol.6 No.-

Sixteen kinds of mushrooms were collected and studied for their agglutinin activities. Among them, several species contained agglutinins(Agaricus bisporus; Rhodophyllus rhodopholius; Marasmius maximus; Pleurotus ostreatus). A lectin component was isolated and partially purified from agglutiinins of Agaricus bisporus. Immunological experiments were performed with this lectin to investigate the properties of other agglutinins.

표고버섯 렉틴의 림프구 자극 분열 및 암 세효 응집 효과

문익재,정시련,전경희 영남대학교 약품개발연구소 1995 영남대학교 약품개발연구소 연구업적집 Vol.5 No.-

A lectin from the edible mushroom. Lentinus edodes, was purified through physiological saline extraction, ammonium sulfate fractionation and column chromatographies. On polyacrylamide gel electrophoresis, 0.05M fraction from hydroxyapatite column exhibited adjacent four sharp bands. The partially purified lectin agglutinated the erythrocytes of rabbit, mouse and rat, but not agglutinated human erythrocytes. The lectin's mitogenic effects were tested by its application to human and murine splenic lymphocytes. The results showed that the 0.05M fraction from hydroxyapatite was mitogenic, and the optimal dose of leninus edodes lectin was slightly lower than Con A by the culture with murine splenic and human peripheral lymphocytes. Meanwhile, its ability to agglutinate transformed cells was tested by its administration to continuous cell lines L1210 and HeLa cells. The lectin was found to be an agglutinin of tumor cell lines teated by L1210 and HeLa cells.

별불가사리(Asteriaa pectinifera) 렉틴의 사이토카인 생성 양상

전경희,최수정,정시련 영남대학교 약품개발연구소 1999 영남대학교 약품개발연구소 연구업적집 Vol.9 No.-

The purpose of this study is to define whether Asterina pectinifera Lectin (APL) is effective on the cytokine production. Isolated mRNA from hPBMC (human peripheral blood mononuclear cells) stimulated with APL for various reaction times (1 to 96 hours) was detected by RT-PCR. The intensity of band for IL-1 and IFNα mRNA was markedly increased at 1 hour, and IL-2 mRNA was strongly expressedat 4 hours. The mRNA band of APL-induced IL-2 and TFNα was weaker than that of IL-1, IL-6 and TNFα. The mRNA expression of 4 cytokines (IL-1, IL-2, IFN?? and TNFα) was detected up to 48 hours, and that of IL-6 was detected until 72 hours. ELISA was used to look protein secretion of the cytokine gene with IL-1, IL-2 and TNFα expressed strongly in RT-PCR. The highest protein secretion was at 4 hours with IL-1, at 8 hours with IL-2 and at 4 hours with TNFα. These results suggest that APL can induce the pro-duction of some cytokines and the immune response from PBMC was done within the first few hours of stimulation with APL.

표고버섯 렉틴의 사이토카인 생성 양상

이인경,김희선,전경희,김성광,정시련 영남대학교 약품개발연구소 1995 영남대학교 약품개발연구소 연구업적집 Vol.5 No.-

The gene expressions of 5 cytokines(IL-1, IL-2, IL-6, TNFα and IFNy) from human peripheral blood mononuclear cells(PBMC) stimulated with Lentinus edodes lectin(LEL) were investigated by using reverse transcription polymerase chain reaction(RT-PCR). PBMC(1×10^(6) cells/ml) isolated from healthy volunteers were stimulated with LEL(O.D.=3.0) or lipopolysaccharide(LPS, 10㎎/ml) for various times(1, 8, 24, 48, 72, 96 and 120 hours). After each of the various stimulated times, total RNA was isolated by RNA sol B method and assessed for various cytokines mRNA by RT-PCR. The mRNA encoding IL-6 and IFNy were detected continuously from 1 to 120 hours. The mRNA encoding IL-1 were detected up to 72 hours and the intensity of the band were increased from 24 to 72 hours. The mRNA encoding IL-2 were detected from 8 to 120 hours and showed delayed reaction compare with other cytokines, But the mRNA encoding TNFα was detected up to 24 hours only. The patterns of TNfα gene expression showed short time response compared with other cytokines. These results suggest that LEL as an inducers of cytokines, can elicits a detectable cytokine on mRNA from the PBMC within the first few hours of stimulation.

Isolation, purification and characterization of phytohemagglutinating proteins from Korean natural products

Chung, See-Ryun,Jeune-Chung, Kyung-Hee,Kim, Kyong-Ae The Pharmaceutical Society of Korea 1980 Archives of Pharmacal Research Vol.3 No.1

Seeds or beans of 55 plants belonging to 31 families were screened by using several different types of red blood cells to find new lectins. In this paper, white kidney been (phaseolus vulgaris C.) was chosen to study biochemical properties of hemagglutinating proteins(lectins). An anion exchanger, DEAE Sephadex A-50, and polyacrylamide disc gel electrophoresis were main techniques used. From three main fractions eluted by stepwise NaCl gradient in 25mM Tris-HCI buffer on DEAE Sephadex column, principal lectin was identified.

Studies on Lectins from Marine Shells (III) : Screening of Lectin-like Agglutinins from Marine Shells

Chung, See-Ryun,Kim, Jang-Hwan,Suh, Young-Ah,Jeunechung, Kyung-Hee The Pharmaceutical Society of Korea 1986 Archives of Pharmacal Research Vol.9 No.4

Forty species of marine shells were collected from Korean coasts and studied extensively for their lectin activities by using erythrocytes of human blooc A, AB, B. O group and rabbit blood. In total, 7 species contained lectines :Neptunea intersculpta, Omphalius nigerrimus and Scapharca subcrenata, blood group nonspecific; Saxidomus purpuratus, human blood A and AB group specific; Lepidozona coreana, Tegilarca granosa and Neptunea polycosta, rabbit blood specific.

Studies on Lectins from Higher Fungi

See Ryun Chung,Kyung Hee Jeune,Moo Kyung Kim 한국생약학회 1989 생약학회지 Vol.20 No.2

The purpose of this research is to develop new lectins, which is extremely useful in the studies of bioscience and medicine. Lectin components were studied extensively from Korean native mushrooms (higher fungi). Lectins are carbohydrate binding proteins and have ability to agglutinate cells. By virtue of this characteristic, lectins have become useful tools in cell membrane study, mitogenic study, immunochemical study and isolation and purification of cells. Recently, a certain lectin is applying even in cancer research and therapy. In the present work, we have concentrated our efforts on collection, identification, and screening of lectin activities of 82 species mushrooms from 32 families. And a few selected mushrooms are further studied and lectins were purified through ion exchange chromatography, gel filtration, hydroxyapatite column and so on. Finally, protein and amino acid contents, carbohydrate analysis, electrophoretic studies, immunochemical studies and other biophysicoc hemical studies were carried out to characterize new lectins from mushrooms.

IMMUNOMODULATING LECTINS FROM MARINE NATURAL PRODUCTS

CHUNG, SEE RYUN,LEE, YUNGCHANG,JEUNE, KYUNG HEE 영남대학교 약품개발연구소 1995 영남대학교 약품개발연구소 연구업적집 Vol.5 No.-

The lectins, a very particular bioactive substance, are isolated and purified from natural products and tried to develop as not only medical uses but also biochemical reagents mainly in advanced country. Since these are macro molecules, obtaining through chemical synthesis is not accomplished yet. For this reason, lectins are extremely expensive. For example, according to the Sigma Chemical Co 1995 catalogue, plant lectins Abrin A costs 59.20$/㎎, Con A type V is 158.20$/g PHA-L is 42.00$/㎎ and a marine animal lectin Limulin costs 125.10$/㎎. This research is designed based on our many years of lectins research experiences throughout the world. We have collected over 100 marine natural products (mainly animals). For these marine natural products, our research team has been tried a series of intensive studies accompanying by modem biochemical techniques. These are lectins screening, extraction, separation, purification of the lectin molecules, purity identification, carbohydrate specificity, heat and pH effects on the activity, determination of molecular size, identification of pI, amino acids assay, carbohydrate assay, immunochemical studies, lymphocytes mitogenic activity lymphokine secretion, cancer cell agglutinability, etc. The four marine animals (Asterina pectinifera, Lunella coronata coreenisis, Meretrix lusoria, and Neptunea intersculpta) turned out as important lectin sources through our recent researches. And continuing further investigations are strongly recommended for the development of new drugs as immunomodulators or as cancer treatment drugs.

Isolation and Purification of New lectins from Leguminosae

Chung, See Ryun,Chung, Kyung Hee Jeune 생화학분자생물학회 1987 BMB Reports Vol.14 No.2

In order to find new lectins from Korean natural products, seeds or beans of 55 kinds plants belonging to 31 families were screened by using several different type of red blood cells. 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 0.1 M 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. A further purification was achieved by hydroxyapatite column chromatography and strong hemagglutinating activity were recovered in 0. 1 M peak. It was electrophoresed and found to migrate as a single band. These suggest that the condition of purif ication for W. K. B. lectin was encourageable but did not brought agreeable satisfaction to the Cercis chinensis lectin even by these very same methods.

미꾸라지 렉틴 성분의 생화학적 특성

Chung, See Ryun,Kim, Jang Hwan,So, Myung Suk,Kim, Moo Kyung,Hyun, Tae Geum,Jeune, Kyung Hee 영남대학교 약품개발연구소 1992 영남대학교 약품개발연구소 연구업적집 Vol.2 No.-

Two kinds of new lectin fractions (LOA-Ⅰ, LOA-Ⅱ) were obtained from loach (Misgurnus spp.) meat by 0.15 M NaCl extraction, salt fractionation, ion exchange and hydroxyapatite column chromatographies. On polyacrylamide gel electrophoresis, LOA-Ⅰ exhibited one major and a few minor bands, but LOA-Ⅱ exhibited three minor bands. The partially purified loach lectins agglutinated not only erythrocytes of human B and AB type, rabbit, dog, but also murine splenic lymphocytes. Agglutinability was relatively labile at various pH and stable at increasing temperature, but was not affected by tested several metal ions. By the sugar specificity test, D-glucosamine and metyl-β-galactopyranose inhibited agglutinating activity at a final concentration of 3 mM. The lectins contained relatively high amounts of aspartic acid, valine and leucine, but sulfur containing amino acids, cystein, methionine and isoleucine were not determined. LOA-Ⅰ, LOA-Ⅱ lectins were nonmitogenic toward murine lymphocytes.