• Archives of Pharmacal Research
• /
• 제9권3호
• /
• pp.119-125
• /
• 1986

Digitonin is a strong hemolysin and glycyrrhizin has protective activity against the deterring effect of other hemolytic saponins. The interaction of these saponins with liposomes was studied as a function of cholesterol in membrane. In the case of multilamellar vesicles, which act as ideal osmometers, digitonin distrupted the barrier function of liposomes composed of phosphatidyl choline, dicetyl phosphate and cholesterol, however, did not influence on cholesterol-lacking liposomes. Glycyrrhizin had similar effect on liposomes irrespective of cholesterol in membrane. In the test with large unilamellar vesicles, digitonin increased the lysis with increasing cholesterol content in membrane, but glycyrrhizin showed no detectable change in cholesterol-containing liposomes. These results suggest that incorporation of cholesterol into liposomes increases the susceptibility to digitonin, resulting in lysis of liposomes, and that the inhibitory effect of glycyrrhizin against other hemolytic saponins in cholesterol-independent.

• The Korean Journal of Physiology
• /
• 제7권2호
• /
• pp.67-70
• /
• 1973

This experiment was carried out to investigate the influence of adrenergic receptor blockade. on digitalis intoxication. The effects of adrenergic alpha and beta receptor blockade on the lethal dose of digitonin were evaluated. $LD_{50}$ and dose mortality curve of digitonin in mice pretreated with dibenzylin or propranolol hydrochloride (Inderal) were obtained. All drugs were injected subcutaneously. Digitonin toxicity was significantly decreased in mice pretreated with beta·blockade compare with alpha-blockade and control groups.

• 대한화학회지
• /
• 제49권5호
• /
• pp.473-478
• /
• 2005

• Journal of Ginseng Research
• /
• 제19권2호
• /
• pp.122-126
• /
• 1995

The effects of treating bovine heart mitochondria with potassium chloride and surfactants such as digitonin and n-dodecy-$\beta$-maltoside (DMS) including plant saponins on extracting cytochrome c were examined. The spectra given by the cytochrome c-containing solutions from the extraction were inspected to ascertain whether ginseng and bellflower saponins could be used instead of the generally- employed surfactants of digitonin and DMS. These studies implied that the effect of ginseng saponins is superior to that of digitonin but inferior to that of DMS, and give rise to the idea of substitutional property of ginseng saponins for the widely-employed surfactants in the extraction of mitochondria intermembrane cytochrome c. The substitution for the solubilizing surfactants by bellflower saponins could, however, not presumably be anticipated; while ginseng saponin mixture are a suitable substitute.

• 한국동물학회지
• /
• 제2권1호
• /
• pp.11-16
• /
• 1959

가물치(Ophice phalus argus)의 안구망막에 존재하는 시각물질을 spectrophotometry 한 결과 다음과 같다. (1) 암순응된 가물치 안구망막에서 2 % digitonin 으로 시각물질을 추출하여 광조사전의 흡광 spectrum을 측정한 결과 흡광 maximum 은 350$\mu\textrm{m}$, 420$\mu\textrm{m}$, 550$\mu\textrm{m}$, 590$\mu\textrm{m}$에 있으며 있었으며 광조사후의 흡광 spectrum과의 difference spectrum 의 흡광 maximum 은 390$\mu\textrm{m}$, 427 $\pm$2$\mu\textrm{m}$, 550$\mu\textrm{m}$, 595$\mu\textrm{m}$에 있었다. (2) 가물치 망막중의 시각물질인 390$\mu\textrm{m}$, 427$\mu\textrm{m}$, 550$\mu\textrm{m}$, 595$\mu\textrm{m}$의 삼원색광에 대한 감광도는 다음과같다. a. 390$\mu\textrm{m}$은 blue light > red light = green light > white light . b.427$\mu\textrm{m}$ 은 red light >blue light > white light > green light c. 550 $\mu\textrm{m}$과 595$\mu\textrm{m}$ 은 blue light > red light > green light > white light (3) 390 $\mu\textrm{m}$ 과 427$\mu\textrm{m}$에서 흡광 spectrum의 maximum을 가지는 시각물질은 그 존재를 예시하던 새로운 색소이다. (4) 가물치의 시각물질은 증류수에도 용출되나 2 % digitonin 수용액으로 추출하였을때보다 명확하지 못하였다. (5) 가물치는 담수어임에도 불구하고 거북류와 같이 visual voilet 는 발견할 수 없었다.

• Parasites, Hosts and Diseases
• /
• 제31권3호
• /
• pp.269-276
• /
• 1993

폐흡충 성충 표피의 구성 단백질의 분자량을 측정하고. 항원성을 갖는 표피 단백질을 규명하였다. 10개월 성충을 0.1% digitonin용액에 담궈 표피하 기저 막까지 박리하여 회수한 표피층을 초음파 분쇄하여 표피 단백질을 추출하였다. 표피 단백 질 추출액을 SDS-PAGE와 immunoblot으로 분석하였다 폐흡충 표피의 구성 단백질은 분자량이 각각 94. 74 (76-66). 62. 54. 44. 42. 38, 28, 26, 25 24. 17. 15.5. 13.5 IEDa으로 측정되었고. 94 kDa 단백질이 가장 주된 표피단백질이었다. 폐흡충 감염자 혈청을 사용한 immunoblot에서 항원성이 확인된 표피단백질의 분자량은 각각 94. 90. 78. 76. 74. 68. 65. 62. 60, 59. 54 kDa 이었다. 이 표피항원 단백질은 폐흡충증 혈청 10개중 7개에서 immunoblot 양성반응을 나타내었고. 7개 양성반응 혈청 각각에서 표피항원 단백질의 특이 항원성이 모두 관찰되었다. 그러나. 이들 폐홉충 표피항원은 간흡충란 양성자혈청과는 전혀 반응하지 않았다. 이상의 결과로. 폐흡충 표피 단백질중에서 분자량 94-54 kDa 사이의 것들이 폐흡충 특이 항원임을 알 수 있었고. 특히 94 kDa 단백질은 가장 양이 많은 대표적인 표피단백질이면서 아울러 특이항원성도 갖고 있었고. 양이 적은 표피단백질이었던 76. 66 kDa 단백질도 상대적으로 높은 특이 항원성을 보여주고 있음을 확인할 수 있었다.

• 생약학회지
• /
• 제6권3호
• /
• pp.125-129
• /
• 1975

The pharmacognostical and pharmacological studies of 'Manryong' were carried out, which has been widely used in Chun-Ra Province, Korea and the following results were obtained. 1) The original plant of the bulb MANRYONG is Erythronium japonicum DECNE. (Liliaceae). 2) The internal structure consists of mostly parenchyma containing numerous starch grains, vascular bundles and substituted fibers. 3) The external surface occurs as a yellowish white and slightly bented ovoidal shape. 4) 'Manryong; contains 11 kinds of free amino acids, such as asparagin, tryptophan, cystine, glutamic acid, threonine, glycine, leucine, proline, histidine, methionine, and alanine. 5) Hemolytic action of the crude saponin isolated from 'Manryong' is weaker than that of digitonin. 6) Effects of 'Manryong' extracts on ceruloplasmin were studied and an antidotal activity of the extracts was found in liver intoxicated with carbon tetrachloride.

• 한국미생물·생명공학회지
• /
• 제23권3호
• /
• pp.311-315
• /
• 1995

Assay conditions for screening of $\beta$-1,3-glucan synthase inhibitor were evaluated. Cells in the beginning of mid-log phase showed the highest activity of the $\beta$-1,3-glucan synthase. Cells permeabilized with 1% digitonin treatment could be used as a good crude enzyme source for convenient screening of the $\beta$-1,3-glucan synthase inhibitors. Calcofluor white (0.125% in final) and papulacandin B (25 $\mu$g/ml) inhibit 90% and more than 50% of the $\beta$-1,3-glucan synthase activity, respectively. Cells grown at 37$\circ$C showed higher enzyme activity than those of 25$\circ$C. Catalytic factor of the $\beta$-1,3-glucan synthase was solubilized from particulated membrane preparations, holoenzyme, by extracting with 0.00938% CHAPS.

• Biomolecules & Therapeutics
• /
• 제1권2호
• /
• pp.188-195
• /
• 1993

In order to understand the machanism of action and regulation of ${\beta}$-adrenergic receptor in terms of molecular level, the purification of receptor protein has a fundamental importance. Moreover, species differences among avian, amphibian and mammalian ${\beta}$-adrenergic receptors make it more important to purify mammalian ${\beta}$-adrenergic receptor. Because ${\beta}$-adrenergic receptor is an integral membrane protein, it must be solubilized from the membrane for the purification. The purpose of the present study was to solubilize and characterize the mammalian $\beta$-adrenergic receptor from guinea pig lung in quantities by more efficient and practical method eventually to purify receptor. Guinea pig lung membrane preparation was solubilized by sequential treatment of buffers containing low and high concentration of digitonin which are 0.2 and 1.2% respectively. About 50% of the total receptor pool was released by this double extraction procedure. The $\beta$-adrenoceptors in the digitonin extract were identified using the ${\beta}$-adrenergic antagonist, (-)-[$^3H$]-dihydroalprenolol ([$^3H$]DHA). The solubilized receptor retained all of the essential characteristics of membrane-bound receptor, namely saturability; stereoselectivity; high affinity to ${\beta}$-adrenergic drugs. For the measurement of soluble receptor activity, Sephadex G-50 chromatography method has been widely used. Inspite of its accuracy and wide acceptance, this technique employed troublesome column work which required long time to assay the activity of receptor. We employed another methods to measure receptor activity. When using 0.5% polyethylenimine pretreated GF/B glass fiber filter, filtration technique could be used to measure soluble receptor activity. This technique enabled us to reduce the total amount of time to assay by a factor of 4 as well as to detect soluble receptor. In the present study, we could establish more efficient and practical solubilization method of mammalian $\beta$-adrenergic receptor. The rapidity and high yield of this solubilization scheme, together with the favorable recovery of the receptor activity, are significant steps toward the ultimate purification of the mammalian $\beta$-adrenergic receptor. The result of this study together with more convenient purification method could provide large amount of purified receptor with ease for various research purposes.

• Bulletin of the Korean Chemical Society
• /
• 제34권3호
• /
• pp.931-935
• /
• 2013

Lipid events in liposome-mediated transfection (lipofection) are largely unknown. Here we studied whether phospholipase D (PLD), an important enzyme responsible for phospholipid breakdown, was affected during lipofection of HepG2 cells with a luciferase plasmid. Synthetic cholesterol (Chol) derivatives, including $3{\beta}$[L-ornithinamide-carbamoyl]Chol, [polyamidoamine-carbamoyl]Chol and $3{\beta}$[N-(N',N'-dimethylaminoethane)-carbamoyl]Chol, and a cationic lipid, N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride were mixed with a helper lipid dioleoylphosphatidylethanolamine to form respective cationic liposomes. All cationic liposomes were found to stimulate PLD. Although orders of magnitude effects of the cationic liposomes on PLD stimulation did not consistently match those on cytotoxicity and luciferase expression, a causal relationship between PLD activation and cytotoxic effect was remarkable. PLD stimulation by the cationic liposomes was likely due to their amphiphilic characters, leading to membrane perturbation, as supported by similar results obtained with other membrane-perturbing chemicals such as oleate, melittin, and digitonin. Our results suggest that lipofection induces cellular lipid changes such as a PLD-driven phospholipid turnover.