• Title/Summary/Keyword: affinity binding

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리포솜-아미노산 결합체의 제조와 포도당 민감성에 대한 연구

  • Mun, Je-Yeong;Lee, Gi-Yeong;Kim, Jin-Cheol;Park, Gi-Nam
    • 한국생물공학회:학술대회논문집
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    • 2000.11a
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    • pp.755-758
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    • 2000
  • Glucose sensitive liposomes-amino acid cinjugates Were prepared by DPPC and asparagine derivatives. Liposomes(amino acid added) have higher glucose binding affinity than liposome(amino acid non-added) or distilled water. The liposomes stabilily were increased by adding cholesterol. Liposomes(cholesterol non-added) particles size were bigger than cholesterol added liposomes.

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Effect of Methanethiol on the Xanthine Oxidase Activity of Liver and Serum in Rats (흰쥐에 Methanethiol 투여가 간 및 혈청 Xanthine Oxidase 활성에 미치는 영향)

  • 윤종국;정소웅;이상일
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.20 no.3
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    • pp.209-213
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    • 1991
  • To evaluate the effect of methanethiol on the activity of xanthine oxidase in both liver and serum, the rats intraperitoneally received methoanethiol. Injection of methoanethiol in rats showed the more decreased xanthine oxidase activity of boty liver and serum than that of the control group. Concomitantly, the xanthine oxidase activity in livers preincubated with methanethiol was decreased in vitro. The xanthine oxidase in livers preincubated with the methoanethiol also showed more increased Km and similar Vmax value than those of the control. These results suggest that the methanethiol may induce a change in substrate binding affinity of xanthine oxidase.

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PHENOTYPING OF PHENYTOIN TREATED RAT LIVER CYTOCHROME P-450 WITH MONOCLONAL ANTIBODIES

  • Jin, Hak-Song;Won, Sun-Me;Park, Sang-Shin;Kim, Nak-Doo
    • Toxicological Research
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    • v.7 no.1
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    • pp.1-12
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    • 1991
  • The phenotyping of cytochrome P-450 in hepatic mivrosomes induced by phenytoin in the rats was carried out by using several monoclonal antibodies (MAbs) against specific P-450 isozymes. Phenytoin (180 mg/kg) was administered intrapritoneally for three consecutive days to the male Sprague-Dawley rats(100-120g). Solid phase radio-immunoassay showed higher binding affinity of MAb PB 2-66-3 and PCN 2-13-1 to the microsomes from phenytoin treated rats than those to from untreated rats, which was comparable to the level in phenobarbital induced rat hepatic microsomes.

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Effect of Testosterone on Central Noradrenergic Nervous System and LHRH (중추 노르아드레날린성 신경계 및 황체호르몬 분비 촉진호르몬에 대한 테스토스테론의 영향)

  • 고홍숙;김경진;박종세;고광호
    • YAKHAK HOEJI
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    • v.35 no.4
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    • pp.295-300
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    • 1991
  • Ralationship between noradrenergic nervous system activity and luteinizing hormone releasing hormone(LHRH) content mediated by testosterone in hypothalamus was tested. Three groups of adult male animals were prepared; (1) Intact; (2) Castration+Vehicle (Cast+V); (3) Castration+Testosterone (Cast+T). Silastic capsule containing vehicle or testosterone was implanted into neck region of animals two weeks following castration. Norepinephrine content, alpha-adrenergic receptor binding characteristics using H$^{3}$-WB4101, and content of LHRH by LHRH RIA procedure were determined. Testosterone replacement to castrated male rats augmented the content of norepinephrine and LHRH. Testosterone replacement increased the alpha-adrenergic receptor density but did not change alpha-receptor affinity. The data from the present study suggest that increase in LHRH content by testosterone may be positively coupled to the activity of central noradrenergic nervous system.

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METALLOTHIONEIN GENE EXPRESSION BY CADMIUM IN CRUCIAN CARP (CARASSIUS AURATUS)

  • Nam, Seong-Sook;Heekyung Bae;Kim, Eunkyoung;Moon, Chang-Kiu;Jeon, Seong-Hwan;Na, Jin-Gyun;Park, Kwangsik
    • Proceedings of the Korean Society of Toxicology Conference
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    • 2002.05a
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    • pp.127-127
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    • 2002
  • Methallothioneins(MTs) are low-molecular-mass cysteine-rich metal-binding proteins with high affinity for heavy metal ions, found in a large variety of organisms. Although the biological functions of MTS have not been fully elucidated, they are thought to play an important role in detoxification of toxic elements such as cadmium and mercury.(omitted)

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In Vitro Selection of RNA Aptamer Specific to Salmonella Typhimurium

  • Han, Seung Ryul;Lee, Seong-Wook
    • Journal of Microbiology and Biotechnology
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    • v.23 no.6
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    • pp.878-884
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    • 2013
  • Salmonella is a major foodborne pathogen that causes a variety of human diseases. Development of ligands directly and specifically binding to the Salmonella will be crucial for the rapid detection of, and thus for efficient protection from, the virulent bacteria. In this study, we identified a RNA aptamer-based ligand that can specifically recognize Salmonella Typhimurium through SELEX technology. To this end, we isolated and characterized an RNase-resistant RNA aptamer that bound to the OmpC protein of Salmonella Typhimurium with high specificity and affinity ($K_d$ ~ 20 nM). Of note, the selected aptamer was found to specifically bind to Salmonella Typhimurium, but neither to Gram-positive bacteria (Staphylococcus aureus) nor to other Gram-negative bacteria (Escherichia coli O157:H7). This was evinced by aptamer-immobilized ELISA and aptamer-linked precipitation experiments. This Salmonella species-specific aptamer could be useful as a diagnostic ligand against pathogen-caused foodborne sickness.

Antifertility Effect of Passive Immunization against Progesterone Monoclonal Antibody in Mice (Progesterone 단일클론항체의 수동면역이 Mouse 수정란의 착상저해에 미치는 효과)

  • 김정우;김종배;정길생;고대환
    • Korean Journal of Animal Reproduction
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    • v.14 no.3
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    • pp.157-164
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    • 1990
  • Anti-progesterone monoclonal antibody injected intraperitonially as a single dose(100$\mu\textrm{g}$) 48hours post coitum(p.c.) almostly blocked pregnancy in ICR mice. The blocking rate of pregnancy in mice treated with antibody were decreased proportionally according to dose of antibody injected ; the rate were 60%, 57% and 17% as the antibody of 10$\mu\textrm{g}$, 50$\mu\textrm{g}$ and 100$\mu\textrm{g}$ were injected respectively. Blood serum progesterone concentration was greatly increased(21 times) after treatment(100$\mu\textrm{g}$), by virtue of high-affinity binding by antibody in circulation of non-pregnant mice in coompared with that of control group at day 10 p.c.. The concentration was about 1.6 times higher in the pregnant mice than in the non-pregnant mice in antibody treated group. In control group, the progesterone concentration was over 7 times higher in the pregnant mice than in non-pregnant mice at day 5 p.c..

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Farnesyl Protein transferase의 분리, 유전자 재조합 및 발현연구

  • 백영진;유권열;박치욱;양철학
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.55-55
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    • 1993
  • Farnesyl Protein transferase(FPT)는 발암유전자 ras의 단백질 산물인 p$^{21}$의 post-translational modification의 첫 단계인 ras-farnesylation에 관여하는 효소로 본 연구에서는 정제된 FPT와 E. coli에서의 발현 system을 이용하여 FPT의 구조와 기능을 밝히고 이를 FPT 방해제의 설계에 이용하고자 한다. Bovine testis에 존재하는 FPT를 30%-50%의 Ammonium sulfate로 fractionation하고, DEAE-Sephacel, Sephacryl S-300 column을 통과시킨 후 peptide(KKCVIM) affinity column을 이용하여 순수 정제하였다. 정제된 효소의 분자량은 gel-filtration에 의해 100KDa으로 추정되었고 SDS-PAGE 결과 49KDa과 46KDa의 두 subunit로 구성되었음이 확인되었다. 효소활성에는 $Mg^{2+}$$Zn^{2+}$가 필수적이며 최적 pH는 7.0이었다. Yeast의 FPT의 두 subunit 유전자는 Yeast genomic DNA를 template로 사용하고 각 subunit에 specific한 합성된 primer들과 vent polymerase를 이용하여 Polymerase chain reaction을 통하여 얻었다. 두 유전자를 pBluescriptII SK+ vector를 변형시킨 두 vector, pBSK+4와 pBChl+4에 재조합 시킨 후 E.coli에 transformation시켜 발현시켰다. 현재 정제된 Bovine FPT와 E. coli에서 발현된 Yeast FPT의 chemical modification과 site-directed mutagenesis를 통하여 FPT의 active site와 substrate binding site에 관한 연구를 진행시키고 있다.

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Quantitative Evaluation of Liver Function with Hepatic Receptor Scintigraphy using Tc-99m Galactosylated Serum Albumin (GSA) (Tc-99m Galactosylated Serum Albumin (GSA)을 이용한 정량적 간수용체 영상술)

  • Lee, Jae-Tae
    • The Korean Journal of Nuclear Medicine
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    • v.32 no.4
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    • pp.305-313
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    • 1998
  • The reduction in the amount of asialoglycoprotein (ASGP) receptor, which resides exclusively on the plasma membrane of functioning mammalian hepatocytes, as a consequence of hepato-cellular damage has been demonstrated in various pathologic conditions of the liver. Galac-tosylated human serum albumin (GSA) is a newly developed receptor-binding agent, specific for the ASGP receptor. Tc-99m GSA binds quantitatively to liver ASGP receptors and the rate of accumulation in the liver is dependent on hepatic function represented as the amount of receptor, as well as the amount of ligand injected, its affinity to the receptor and the hepatic blood flow. The findings of Tc-99m GSA scintigraphy were reported to reflect the hepatic function of the patients with large hepatic tumors, obstructive jaundice, acute and chronic liver disease. Tc-99m GSA scintigraphy is an easy and reliable test and has the clinical potentials to evaluate the liver function in the patients with hepatic disorders.

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Analysis of In Vivo Interaction of HCV NS3 Protein and Specific RNA Aptamer with Yeast Three-Hybrid System

  • HWANG BYOUNGHOON;LEE SEONG-WOOK
    • Journal of Microbiology and Biotechnology
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    • v.15 no.3
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    • pp.660-664
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    • 2005
  • We have previously isolated specific RNA aptamers with high affinity against the helicase domain of hepatitis C virus (HCV) nonstructural protein 3 (NS3). The RNA aptamers competitively and efficiently inhibited the helicase activity, partially impeding HCV replicon replication in human hepatocarcinoma cells. In this study, the RNA aptamers were tested for binding to the HCV NS3 proteins in eukaryotic cells, using a yeast three-hybrid system. The aptamers were then recognized by the HCV NS3 proteins when expressed in the cells, while the antisense sequences of the aptamers were not. These results suggest that the in vitro selected RNA aptamers can also specifically bind to the target proteins in vivo. Consequently, they could be potentially utilized as anti-HCV lead compounds.