• 제목/요약/키워드: concanavalin A

검색결과 211건 처리시간 0.027초

Purification of the Glycosylated Polyphenol Oxidase from Potato Tuber

  • Kwon, Do-Yoon;Kim, Woo-Yeon
    • BMB Reports
    • /
    • 제29권2호
    • /
    • pp.163-168
    • /
    • 1996
  • Glycosylated polyphenol oxidase was purified from potato tuber using ammonium sulfate fractionation, Sephadex G-100, and concanavalin A Sepharose column chromatography. Two or three types of polyphenol oxidase were separated on concanavalin A Sepharose. Type I and II polyphenol oxidases did not bind to concanavalin A Sepharose. Type I seemed to be an aggregated form of polyphenol oxidase. Type III polyphenol oxidase, which is presumed to be glycosylated because it was bound to concanavalin A Sepharose and eluted with $\alpha$-D-methyl glucopyranoside, was further purified by chromatography on Econo-Pac Q and Superose 12. Glycosylated polyphenol oxidase was purified 130-fold from the dissolved ammonium sulfate pellet resulting in about $6\;{\mu}g$ of the enzyme from 100 g of potato tuber periderm. The molecular weight of the glycosylated enzyme determined by SDS-polyacrylamide gel electrophoresis was about 64,000. Optimum temperature and pH of both II and type III potato polyphenol oxidases were $20^{\circ}C$ and pH 7.0, respectively. Glycosylated form of polyphenol oxidase (type III) preferred catechol to catechin as a substrate, whereas type II enzyme showed the reverse substrate preference.

  • PDF

근소포체의 단백질 및 당단백질 조성에 관한 연구 (Studies on the Compositon of Protein and lycoprotein in Sarcopiasmic Reticulum of Skeletal Muscle)

  • 박영철
    • 한국동물학회지
    • /
    • 제33권2호
    • /
    • pp.191-199
    • /
    • 1990
  • 토끼의 골격근에서 근소포체를 순수 분리하여 SDS-polyacrylamide gel전기영동법과 125 I-concanavalin A표지법으로 단백질과 당단백질의 조성을 조사하였다. 전기영동사에 나타난 대표적인 단백질은 $Ca^2$+-AThase, 80 Kd protein,calsequestrin,high affinity calcium binding protein, intrinsic glycoprotein이었으며, 160 Kd protein, 94 Kd protein,38 Kd protein, 34 Kd protein,24 Kd proteins도 존재하였다.특히, 막성계에 있는 heak protein으로 알려져 있는 80 Kd protein은 본 연구를 통해 주로 근소포체의 terminal cisternae에 들어 있음이 확인되었다. 한편 125 I-concanavalin A표지에 의해 전기영동성에 나타난 대표적인 당단백질은 160 Kd glycoprotein, 94 Kd glycoprotein, calsequestrin, intrinsic glycoprotein의 4종이었다. 이 가운데 94 Kd glycoprotein은 94 Kd glucose-regulated protein으로 추정되며, 본 연구를 통해 근소포체에서도 특히 T-tubule에 다량으로 존재함이 밝혀졌다.

  • PDF

Studies on the Preparation of Radioactive iodine Labelled Concanavalin-A, Lectin Extracted from Korean Native Plant “Banha”, and Their Conjugation Products and the Hemagglutination Tests of These Labelled Compounds in Vitro.

  • Kim, You-Sun;Park, Kyung-Bae
    • Nuclear Engineering and Technology
    • /
    • 제10권1호
    • /
    • pp.1-12
    • /
    • 1978
  • Concanavalin-A, 한국산 반하(半夏)로부터 주출된 Lectin 및 이들과 tyrosine 또는 5-iodo-6-aminouracil을 결합시킨 화합물들을 각각 방사성 요오드-125로 표지 하였으며 표지된 화합물들을 사용하여 생체외부 시험법으로 혈액응고 시험을 실시하였다. 표지반응에 관하여 표지수율과 함께 그 실험조건을 보고하였으며 Lectin과 여러 아미노산계통 화합물들과의 결합반응성을 논의하였다. 암세포에 대한 혈액응고 시험결과를 보고하였으며 이들 표지화합물들의 임상적 실용전망에 관하여서도 논의하였다.

  • PDF

Concanavalin A를 매개로한 세포내 Calcium의 변화와 생쥐 포배의 팽창과 부화 (Concanavalin A Mediated Calcium Changes on Expansion and Hatching of the Mouse Blastocyst)

  • 전용필
    • Clinical and Experimental Reproductive Medicine
    • /
    • 제32권4호
    • /
    • pp.337-346
    • /
    • 2005
  • Objective: The oligosaccharide moieties of glycoproteins and proteoglycans have a vital function in blastocyst differentiation. Concanavalin (ConA), a lectin, is known to bind on the preimplantation embryos, especially on blastocyst. In this study, we investigated whether ConA can modulate the trophoblast development and about the regulating mediator. Also, we investigated whether expansion is enough for hatching procession of the mouse blastocyst. Method: Embryos were collected at 72 h post hCG injection and chemicals were treated after 24 h (96 hr post hCG injection). ConA or calcium ionophore A23187 were exposed to blastocyst and than analysis the developmental process for 48 hr. Intracellular free-$Ca^{2+}$ concentration in trophectoderm was measured with confocal laser microscope after exposing to ConA or calcium ionophore A23187. ConA-pretreated blastocyst exposed to the calcium ionophore A23187 and then analyzed the developmental process. Otherwise ouabain was treated to the blastocyst to block the $Na^+/K^+$-ATPase activity. Results: In contrast to the control blastocyst, the ConA-exposed blastocysts developed beyond the expansion stage with significantly high rate (90.4%) at 12 h post administration. ConA induced an increase the intracellular $Ca^{2+}$ concentration in trophectoderm. Calcium ionophore A23187 also stimulated expansion of blastocyst. Most of the control blastocysts developed to the hatching stage at 144 h post hCG injection. However, strongly 65% of the ConA-exposed embryos were arrested at expanded stage at same time point. The developmental progression rates to hatching stage of both ConA- and calcium ionophore A23187-expose blastocysts were significantly lower than that of the control. However ConA-pretreated embryos developed to the hatching stage like control embryos. Ouabain showed a tendency to delayed the progress to expansion stage but did not inhibit the development to the hatching stage. Conclusion: ConA-mediated expansion is the result of the increase of intracellular free-calcium in blastocyst stage embryo. It is suspected that expansion of the blasocyst is a essential indirect factor in hatching and the calcium may triggering the cellular mechanisms for the both expansion and hatching progression.

Heterometal-Coordinated Monomeric Concanavalin A at pH 7.5 from Canavalia ensiformis

  • Chung, Nam-Jin;Park, Yeo Reum;Lee, Dong-Heon;Oh, Sun-Young;Park, Jung Hee;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
    • /
    • 제27권12호
    • /
    • pp.2241-2244
    • /
    • 2017
  • The structure of concanavalin A (ConA) has been studied intensively owing to its specific interactions with carbohydrates and its heterometal ($Ca^{2+}$ and $Mn^{2+}$) coordination. Most structures from X-ray crystallography have shown ConA as a dimer or tetramer, because the complex formation requires specific crystallization conditions. Here, we reported the monomeric structure of ConA with a resolution of $1.6{\AA}$, which revealed that metal coordination could trigger sugar-binding ability. The calcium coordination residue, Asn14, changed the orientation of carbohydrate-binding residues and biophysical details, including structural information, providing valuable clues for the development and application of detection kits using ConA.

Comparison of Lectin from Pseudixus japonicus and Concanavalin a on Lymphocytes Proliferation and Cytotoxicity

  • Chung, Yong-Za;Jung, Hyun-Ok;Hong, Tae-Hong;Suh, Sok-Soo
    • Archives of Pharmacal Research
    • /
    • 제14권3호
    • /
    • pp.207-216
    • /
    • 1991
  • Pseudixus japonicus agglutinin (PJA) was isolated. And its characteristics were compared with those of concanavalin A (Con A). PJA is a glycopritein composed of 49.3% carbohydrate and 50.7% protein which had relatively high percentages of glutamic acid, aspartic acid and phenylalanine residues. The hemagglutinating activity of PJA was approximately one-eighth of that of Con A when tested with mouse crythrocytes. PJA failed to simulate the proliferation or transformation of human and mouse lymphocytes in contratst to Con A. PJA and Con A showed cytotoxicities against SNU-1 (human stomach cancer cells), SNU-CI (human colon cancer cells) and mouse Sarcoma 180 cells when tested by 3-(4, 5-dimethyl thiazol-2-yl)2. 5-diphenyl tetrazolium bromide (MIT) colorimetric assay. The antitumor activity of the lectin in vivo was also tested in Sarcoma 180 bearing mice. There was no significant difference in prologation of lifc span of the mice after the treatment with PJA and Con A for 10 consecutive days.

  • PDF

T-2 Toxin이 병아리 비장세포의 유전질 발생에 미치는 영향 (Effect of T-2 Toxin on the Mitogen-Induced Blastogenesis in Chick Splenic Cell)

  • 전향숙;정덕화;이서래
    • 한국식품과학회지
    • /
    • 제26권5호
    • /
    • pp.585-589
    • /
    • 1994
  • 시험관 내에서 T-2 toxin이 병아리 비장세포의 blastogenesis에 미치는 영향을 살펴본 결과, B-cell mitogen인 lipopolysaccharide 및 T-cell mitogen인 concanavalin A 자극에 대해 T-2 toxin의 농도가 증가함에 따라 억제정도가 증가하는 경향을 나타내었다. 노출시기를 달리하여 T-2 toxin을 투여한 병아리의 비장세포에서 mitogen 자극에 내한 반응을 안아 본 결과, 부화하기 전과 후에 계속 T-2 toxin에 노출시킨 실험군은 가장 영향을 많이 받은 것으로 나타났고 부화전 혹은 부화후 어느 한 시기에만 T-2 toxin에 노출된 실험군은 비교적 영향을 적게 받는 것으로 나타났다.

  • PDF

Lupane계 Triterpenoid류가 임파구 분열에 미치는 효과 (Effects of Lupane-Triterpenoids on Mitogen-induced Proliferation of Lymphocytes)

  • 김영옥;조대현;정혜주;김진호;장승엽;육창수;양기숙;오오진
    • 약학회지
    • /
    • 제43권2호
    • /
    • pp.208-213
    • /
    • 1999
  • The effects of five lupane-triterpenoids from leaves of two Acanthopanax spp., chiisanogenin, chisanoside and $22{\alpha}-hydroxychiisanogenin$, acakoreoside A and acantrifoside A on the mitogen-induced proliferation were investigated in vitro. T cell proliferation (TCP) to concanavalin A (Con A) and the B cell proliferation (BCP) to lipopolysaccharide (LPS) were increased by chiisanogenin. TCP to Con A was significantly increased by chiisanoside and acankoreoside A, but not affected by chiisanogenin, $22{\alpha}-hydroxychiisanogenin$ and acantrifoside A, BCP to LPS was significantly increased by acankoreoside A and acantrifoside A, and slightly increased by chiisanoside, chiisanogenin and $22{\alpha}-hydroxychiisanogenin$.

  • PDF

Partial Purification and Characterization of a Glycoprotein Factor from Fresh Ginseng

  • Kong, Yun-Cheung;Fong, Wing-Ping;Song, Myung-Eun;Ng, Kam-Hung;Ho, Dan-Dan;Ng, Ping-Chung
    • 고려인삼학회:학술대회논문집
    • /
    • 고려인삼학회 1990년도 Proceedings of International Symposium on Korean Ginseng, 1990, Seoul, Korea
    • /
    • pp.79-85
    • /
    • 1990
  • The aqueous extract of fresh ginseng (Panax ginseng C.A. Meyer) contains a macromolecular fraction that showed mitogenic and comitogenic activities in human peripheral blood Iymphocytes. Purification of the crude extract by size (ultrafiltration, Sephadex G-200) and charge (DEAE-cellulose. DEAE-Sepharose) yielded a semi-purified fraction (DS-3). This fraction contains at least three subgroups of anionic macromolecules with apparent molecular weight greater than 600 kilodaltons. It is a glycoprotein with a large amount of glucuronic acid. It acts as a mitogen in both T and B cells of human peripheral blood lymphocytes. It could also potentiate the mitogenic action of Concanavalin A in Iymphocyte T cells. Such potentiation is not due to increased binding of Concanavalin A to the cell surface. Its mitogenic and co-mitogenic effects do depend on the presence of extracellular Ca2+.

  • PDF