• 제목/요약/키워드: affinity interaction

검색결과 240건 처리시간 0.025초

Specific Targeting of Fluorescein Isothiocyanate with Ep-CAM Antibody(Specific targeting of FITC with Ep-CAM Antibody)

  • Lee, Young-Tae;Tae, Gun-Sik
    • Journal of Photoscience
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    • 제10권3호
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    • pp.237-240
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    • 2003
  • The tetradecameric peptide (K47-K60) near the NH$_2$-terminal region of epithelial-cell adhesion molecule (Ep-CAM) was chosen as antigenic site and a polyclonal antibody was generated, which could recognize Ep-CAM from the mouse colon tissue or the colon cancer cell, CT-26, in Western blot analysis. Then, the fluorescein isothiocyanate (FITC), a fluorescence dye, was conjugated with the affinity purified Ep-CAM antibody using thiocyanate and the amino groups of FITC and antibody, respectively. The molar ratio of FITC to antibody was estimated approximately 1.86 to 1.00 by measuring the optical densities at 492 nm and 280 nm. Ep-CAM antibody-FITC conjugate was then used for immunohistochemistry of the CT-26 cells. Judging from the shapes formed by fluorescence, the Ep-CAM antibody could delivered FITC to the surface of cells in which Ep-CAM was expressed. This result implies that Ep-CAM antibody could be also used for the tissue-specific delivery of the photosensitizer to the target protein via antigen-antibody interaction.

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대장균 리보스 결합단백질의 신호배열 변이에 대한 숙성체 부위의 회복돌연변이 (Intragenic Suppressors for Expory-defective Signal Sequence Mutation of Ribose-binding Protein in Escherichia coli)

  • 이영희;송택선;김정호;박순희;박찬규
    • 미생물학회지
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    • 제29권5호
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    • pp.270-277
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    • 1991
  • A mutational alteration in the signal sequence of ribose-binding protein (RBP) of Escherichia coli, rbsB103, completely blocks the export of the protein to the periplasm. Intragenic suppressors for this mutation have been selected on minimal medium with ribose as a sole carbon source. Six suppressor mutations were characterized in detail and were found to have single amino acid wubstitution in the mature portion of RBP, which resulted in the mobility shift of the proteins on SDS polyacrylamide gel. Amino acid changes of these suppressors were localized in several peptides which are packed to form the N terminal domain of typical bilobate conformation of RBP. The involvement of SecB, a molecular chaperone, was investigated in the suppression of signal sequence mutation. Translocation efficency was found to be increased by the presence of SecB for all suppressors. It is likely that the folding characteristics of RBP altered by the suppressor mutations affect the affinity of interaction between SecB and RBP.

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말벌 독성 물질과 그 유도체의 인지질 막 환경에서의 상호작용 (Interaction of Hornet Venom and its Derivatives in the Phospholipid Membrane Environment)

  • 이봉헌;박홍재
    • 한국환경과학회지
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    • 제7권1호
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    • pp.62-66
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    • 1998
  • Toxic Mastoparan B(MP-B) which is purified from the venom of the hornet Vespa basalis is a cationic amphlphilic tetradecapeptide. MP-B and Its Ala-substituted analogues were synthesized by solld phase method and the toxic peptide-membrane interactions were examined by circular dichroism(CD) spectra, fluorescence spectra, and leakage abilities in phospholipid membranes. In the presence of phospholipid vesicles, synthetic MP-B and its analogues formed amphiphilic -helical structures, but in the buffer soletion, those exhibited random coil conformation as measured by CD. Fluorescence spectra of MP-B and its analogues which indicated the binding affinity of peptide on phospholipid vesicles showed that the replacement of Lys at position 2 and 11 with Ala caused a remarkable effect in the blue shalt and that at position 2, in the leakage ability of the peptide.

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Backbone Assignment of Phosphorylated Cytoplasmic Domain B of Mannitol Transporter IIMtl in Thermoanaerobacter Tengcongensis

  • Lee, Ko On;Suh, Jeong-Yong
    • 한국자기공명학회논문지
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    • 제21권1호
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    • pp.20-25
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    • 2017
  • The cytoplasmic domains A and B of the mannitol transporter enzyme $II^{Mtl}$ are covalently linked in Escherichia coli, but separately expressed in Thermoanaerobacter Tengcongensis. The phosphorylation of domain B ($TtIIB^{Mtl}$) substantially increases the binding affinity to the domain A ($TtIIA^{Mtl}$) in T. Tengcongensis. To understand the structural basis of the enhanced domain-domain interaction by protein phosphorylation, we obtained NMR backbone assignments of the phospho-$TtIIB^{Mtl}$ using a standard suite of triple resonance experiments. Our results will be useful to monitor chemical shift changes at the active site of phosphorylation and the binding interfaces.

복진정제 및 수면제 유발 사건수면 (Sedative Hypnotics Induced Parasomnias)

  • 이유진
    • 수면정신생리
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    • 제19권1호
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    • pp.18-21
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    • 2012
  • Parasomnias induced by hypnosedatives are rare but serious side effect. Such parasomnias have not been reported with all hypnosedatives. However, frequent use of hypnosedatives, particularly nonbenzodiazepine receptor agonists is associated with parasomnias. Associated symptoms are sleep eating, sleepwalking with object manipulation, sleep conversations, sleep driving, sleep sex and sleep shopping etc. Mechanisms include high affinity for $GABA_A$ receptor, interruption of the consolidation phase of memory formation by drug, pharmacokinetic or pharmacodynamic drug-drug interaction and concomitant administration with alcohol. Managements for parasomnias induced by hypnosedatives involve stopping medication, switch to other medications or nonpharmacological treatment, lowest effective dose of NBRAs (Non-Benzodiazepine Receptor Agonists), taking into consideration drug-drug interactions, identification and treatment of underlying disease states.

Purification and Characterization of Glyoxalase I from Chlamydomonas reinhardtii

  • Hwang, Sun-Jun;Chai, Young-Gyu
    • BMB Reports
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    • 제29권4호
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    • pp.294-299
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    • 1996
  • Glyoxalase I (Ee 4.4.1.5, lactoylglutathione lyase) from Chlamydomonas reinhardtii was purified to homogeneity by ammonium sulfate fractionation, anion-exchange chromatography, hydrophobic interaction chromatography, and affinity chromatography on S-hexylglutathione agarose. The purified enzyme was judged to be homogeneous on SDS-PAGE, and consisted of a single polypeptide chain with a relative molecular weight of 24,000. The enzyme was most active at $40^{\circ}C$ and pH 7.5. It was catalytically most active with methylglyoxal as substrate. A number of properties of the Chlamydomonas glyoxalase I enzyme, such as substrate specificity, molecular mass, kinetic parameters, pi, metal ion effect, have been determined and compared with those reported for preparations from other sources. It had somewhat different characteristics from mammalian enzymes.

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Coexpression and protein-protein complexing of DIX domains of human Dvl1 and Axin1 protein

  • Choi, Seung-Hye;Choi, Kyung-Mi;Ahn, Hyung-Jun
    • BMB Reports
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    • 제43권9호
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    • pp.609-613
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    • 2010
  • The Dvl and Axin proteins, which are involved in the Wnt signaling pathway, each contain a conserved DIX domain in their sequences. The DIX domain mediates interaction between Dvl and Axin, which together play an important role in signal transduction. However, the extremely low production of DIX domain fragments in E. coli has prevented more widespread functional and structural studies. In this study, we demonstrate that the DIX domains of Dvl and Axin are expressed noticeably in a multi-cistronic system but not in a mono-cistronic system. Formation of the $DIX_{Dvl1}-DIX_{Axin1}$ complex was investigated by affinity chromatography, SEC and crystallization studies. Unstable DIX domains were stabilized by complexing with counterpart DIX domains. The results of the preliminary crystallization and diffraction of the $DIX_{Dvl1}-DIX_{Axin1}$ complex may prove useful for further crystallographic studies.

Dyeability of photooxidized meta-aramid fabrics to cationic dyes

  • Dong, Yuanyuan;Jang, Jin-Ho
    • 한국염색가공학회:학술대회논문집
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    • 한국염색가공학회 2009년도 제41차 학술발표회
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    • pp.79-80
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    • 2009
  • Aramid fabrics were photooxidized by UV/$O_3$ irradiation. The UV irradiation caused the oxygen content of the aramid fabric surface to increase and reduced the surface zeta potential, coupled with improved water wettability. The modified aramid fabrics showed higher affinity to cationic dyes which may be resultd from the newly introduced electrostatic interaction between cationic dyes and anionic dyeing sites on the photooxidized surface layers. Although color fastness to both staining and rubbing were good to excellent, color fastness to shade change was moderate.

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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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    • 제15권3호
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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.

Preparation of Mucoadhesive Chitosan-Poly(Acrylic acid) Microspheres by Interpolymer Complexation and Solvent Evaporation Method II

  • Cho, Sang-Min;Choi, Hoo-Kyun
    • Archives of Pharmacal Research
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    • 제28권5호
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    • pp.612-618
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    • 2005
  • A mucoadhesive microsphere was prepared by an interpolymer complexation and solvent evaporation method, using chitosan and poly(acrylic acid) (PAA), to prolong the gastric resid ence time of the delivery system. The Fourier transform infrared results showed that microspheres were formed by an electrostatic interaction between the carboxyl groups of the PAA and the amine groups of the chitosan. X-ray diffraction and differential scanning calorimetry analysis showed that the enrofloxacin in the chitosan-PAA microsphere was molecularly dispersed in an amorphous state. Scanning electron microscopy of the surface and the quantity of mucin attached to the microspheres indicated that chitosan-PAA microspheres had a higher affinity for mucin than those of chitosan alone. The swelling and dissolution of the chitosan-PAA microspheres were found to be dependent on the pH of the medium. The rate of enrofloxacin released from the chitosan-PAA microspheres was slower at higher pH; therefore, based on their mucoadhesive properties and morphology, the chitosan-PAA microspheres can be used as a mucoadhesive oral drug delivery system.