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.

Lectins from Marine Invertebrates : A Search for New Drugs Useful in Life Science

Chung, See Ryun,Jeune-Chung, Kyung Hee 한국균학회 1987 韓國菌學會誌 Vol.15 No.3

Lectin(from the Latin legere: to choose) is a sugar binding protein or glycoprotein of non-immune origin which agglutinates cells or precipitates glycoconjugates(1). Cell agglutination is a consequence of the formtion of numerous lectin bridges between surface oligosaccharide determinants on neighbouring cells. Lectins exert a variety of remarkable biologic effects on cells: agglutination of erythrocytes and other types of cell, mitogenic stimulation of lymphocytes, generation of suppressor cells, mediation of Killing of target cells by lymphocytes and macrophages, enhancement of phagocytosis of yeasts and bacteria by macrophages, insulin-like effects on fat cells, toxicity to cells and animals, inhibition of growth and migration of tumor cells, induction of vacuole formation in macrophages, immunosuppressive effects in vivo, promotion of cell adhesion and spreading, induction of platelet release reaction, induction of histamine release from basophils and mast cells, induction of patching and capping of receptors on cell surfaces, peroxide release from macrophages, inhibition of endocytosis of lysosomal enzymes by cultured fibroblasts, inhibition of fungal growth, inhibition of fertilization of ovum by sperm, effect on aplastic anemia, anticytotoxic effect on nitrogen mustard and etc(2, 3). By virtue of these characteristics, lectins have become powerful tools in biochemical and medical researches. During the past decade, lectins have been found in a wide variety of plants, animals and microorganisms(4-7). Studies of the functions of lectins in plants have been reviewed by many authors(8-11). But research on animal lectins has only recently begun; consequently there are only limitted reviews(12-14) and moreover little is known about the lectins from marine animals(15-22). During the past several years, we have concentrated our efforts on the isolation, purification and characterization of new lectins from marine invertebrates(23-29). And through these works, we could anticipate the possibility of application of lectins in life science, particularly in cancer research.

STUDIES ON BIOACTIVE MARINE NATURAL SUBSTANCE, LECTINS : MARINE ANIMAL LECTINS (XII)

Chung, See Ryun,So, Myung Suk,Jeune, Kyung Hee 영남대학교 약품개발연구소 1992 영남대학교 약품개발연구소 연구업적집 Vol.2 No.-

The objective of this research is to find new biopotential substances from marine natural products. 60 species from 22 families of marine mollusks (shells) and 16 species from 11 families of marine invertebrates (in total 76 species of animals from 33 families) were collected and studied extensively for their lectin components. Among these, 14 species from shells and 3 species from invertebrates (17 species in total) turned out as containing lectins components relatively high amounts in their tissue or in body fluid. In order to obtain new lectins, many attempts have been performed from the meat and/or body fluid of animals (i. e., Chlorostoma argyrostoma lischkei, Meretricx lusoria, Neptunea intersculpta, Styela plicata, Tapes philippinarum, etc.) which contained relatively high amounts of lectins. Lectins are obtained from the animals by physical saline extraction, ammonium sulfate salt fractionation, centrifugation, ion exchange chromatography, hydroxyapatite column, chromatofocusing, gel filtration, affinity chromatography and so on. To characterize biophysicochemical properties of this particular components, protein assay, polyacrylamide disc gel and SDS electrophoresis, carbohydrate analysis, carbohydrate specificities, mitogenicity and immunochemical analysis were performed.

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.

Differential Cytokinins Binding to Dioecious Plant Ribosomes

Chung, See Ryun,DURAND, Bernard 생화학분자생물학회 1986 BMB Reports Vol.12 No.4

It has been reported that the administration of auxins, gibberellins and cytokinins to plants affects nuclear transcription. Other observations suggest that cytokinins also have an effect at the translational level. Based on our knowledge of sexual differentiation of the dioecious plant Mercurialis annua, we believe that genetically male meristem is a specific target organ of cytokinins. Exogenous cytokinins provoked obvious changes of these male meristems, including the appearance of certain tRNA species and enzymes specific to female flowers. In the present study, the binding of benzylaminopurine, furfurylaminopurine, dimethylallylaminopurine and zeatin, as well as their less acaive ribosides, were compared in order to detect any relations between physiological action and binding efficiency.

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.

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.