• Title/Summary/Keyword: binding agent

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Crystal Structures of 6-Phosphogluconate Dehydrogenase from Corynebacterium glutamicum

  • Hyeonjeong Yu;Jiyeon Hong;Jihye Seok;Young-Bae Seu;Il-Kwon Kim;Kyung-Jin Kim
    • Journal of Microbiology and Biotechnology
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    • v.33 no.10
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    • pp.1361-1369
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    • 2023
  • Corynebacterium glutamicum (C. glutamicum) has been considered a very important and meaningful industrial microorganism for the production of amino acids worldwide. To produce amino acids, cells require nicotinamide adenine dinucleotide phosphate (NADPH), which is a biological reducing agent. The pentose phosphate pathway (PPP) can supply NADPH in cells via the 6-phosphogluconate dehydrogenase (6PGD) enzyme, which is an oxidoreductase that converts 6-phosphogluconate (6PG) to ribulose 5-phosphate (Ru5P), to produce NADPH. In this study, we identified the crystal structure of 6PGD_apo and 6PGD_NADP from C. glutamicum ATCC 13032 (Cg6PGD) and reported our biological research based on this structure. We identified the substrate binding site and co-factor binding site of Cg6PGD, which are crucial for understanding this enzyme. Based on the findings of our research, Cg6PGD is expected to be used as a NADPH resource in the food industry and as a drug target in the pharmaceutical industry.

The Efficacy of Saururus chinensis on Cervical Cancer Cells : The Inhibitory Effect on the Function of E6 and E7 Oncogenes of HPV Type 16 (삼백초 추출물의 자궁경부암세포 억제 효능)

  • Chung, Yeon-Gu;Lee, Hae-Sook;Lee, Kyung-Ae;Joung, Ok;Oh, Won-Keun;Kim, Kwang-Dong;Lim, Jong-Seok;Moon, Ja-Young;Cho, Yong-Kweon;Park, Sue-Nie;Yoon, Do-Young
    • YAKHAK HOEJI
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    • v.46 no.6
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    • pp.426-432
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    • 2002
  • Cervical cancer is one of the leading causes of female death from cancer worldwide with about 500,000 deaths per year. A strong association between certain human papilloma viruses (HPV type 16 and 18) and cervical cancer has been well known. An extract of Saururus chinensis, named as PE-46, has been used to investigate whether this agent has the ability of inhibiting the oncogenes E6 and E7 of HPV type 16. PE-46 inhibited the proliferation of human cervical cancer cell lines in a dose response manner. PE-46 also inhibited the in vitro binding of E6 and E6AP which are essential for the binding and degradation of the tumor suppressor p53. In addition, PE-46 inhibited the in vitro binding of E7 and Rb which is essential tumor suppressor for the control of cell cycle. The levels of mRNA for E6 and E7 were also decreased by PE-46. SiHa cells treated with PE-46 induced G0/G1 arrest, resulting in inhibition of growth. Our study showed that the PE-46 can inhibit the cervical carcinomas via both inhibition of bindings between oncogenes and tumor suppressors, and inhibition of G1longrightarrowS transition. PE-46 inhibited the oncogenecity of E6 and E7 of HPV 16 type, thus could be used as a putative modulating agent for the treatment of cervical carcinomas caused by HPV.

Assessment of Anti-nutritive Activity of Tannins in Tea By-products Based on In vitro Rumen Fermentation

  • Kondo, Makoto;Hirano, Yoshiaki;Ikai, Noriyuki;Kita, Kazumi;Jayanegara, Anuraga;Yokota, Hiro-Omi
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.11
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    • pp.1571-1576
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    • 2014
  • Nutritive values of green and black tea by-products and anti-nutritive activity of their tannins were evaluated in an in vitro rumen fermentation using various molecular weights of polyethylene glycols (PEG), polyvinyl pyrrolidone (PVP) and polyvinyl polypyrrolidone as tannin-binding agents. Significant improvement in gas production by addition of PEG4000, 6000 and 20000 and PVP was observed only from black tea by-product, but not from green tea by-product. All tannin binding agents increased $NH_3$-N concentration from both green and black tea by-products in the fermentation medium, and the PEG6000 and 20000 showed relatively higher improvement in the $NH_3$-N concentration. The PEG6000 and 20000 also improved in vitro organic matter digestibility and metabolizable energy contents of both tea by-products. It was concluded that high molecular PEG would be suitable to assess the suppressive activity of tannins in tea by-products by in vitro fermentation. Higher responses to gas production and $NH_3$-N concentration from black tea by-product than green tea by-product due to PEG indicate that tannins in black tea by-product could suppress rumen fermentation more strongly than that in green tea by-product.

Inhibition of Adipocyte Differentiation by Anthocyanins Isolated from the Fruit of Vitis coignetiae Pulliat is Associated with the Activation of AMPK Signaling Pathway

  • Han, Min Ho;Kim, Hong Jae;Jeong, Jin-Woo;Park, Cheol;Kim, Byung Woo;Choi, Yung Hyun
    • Toxicological Research
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    • v.34 no.1
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    • pp.13-21
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    • 2018
  • Anthocyanins are naturally occurring water-soluble polyphenolic pigments in plants that have been shown to protect against cardiovascular diseases, and certain cancers, as well as other chronic human disorders. However, the anti-obesity effects of anthocyanins are not fully understood. In this study, we investigated the effects of anthocyanins isolated from the fruit of Vitis coignetiae Pulliat on the adipogenesis of 3T3-L1 preadipocytes. Our data indicated that anthocyanins attenuated the terminal differentiation of 3T3-L1 preadipocytes, as confirmed by a decrease in the number of lipid droplets, lipid content, and triglyceride production. During this process, anthocyanins effectively enhanced the activation of the AMP-activated protein kinase (AMPK); however, this phenomenon was inhibited by the co-treatment of compound C, an inhibitor of AMPK. Anthocyanins also inhibited the expression of adipogenic transcription factors, including peroxisome proliferator-activated receptor-${\gamma}$, CCAAT/enhancer-binding protein a and b, and sterol regulatory element-binding protein-1c. In addition, anthocyanins were found to potently inhibit the expression of adipocyte-specific genes, including adipocyte fatty acid-binding protein, leptin, and fatty acid synthase. These results indicate that anthocyanins have potent anti-obesity effects due to the inhibition of adipocyte differentiation and adipogenesis, and thus may have applications as a potential source for an anti-obesity functional food agent.

Molecular and biochemical characterization of a novel actin bundling protein in Acanthamoeba

  • Alafag Joanna It-itan;Moon Eun-Kyung;Hong Yeon-Chul;Chung Dong-Il;Kong Hyun-Hee
    • Parasites, Hosts and Diseases
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    • v.44 no.4 s.140
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    • pp.331-341
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    • 2006
  • Actin binding proteins play key roles in cell structure and movement particularly as regulators of the assembly, stability and localization of actin filaments in the cytoplasm. In the present study, a cDNA clone encoding an actin bundling protein named as AhABP was isolated from Acanthamoeba healyi, a causative agent of granulomatous amebic encephalitis. This clone exhibited high similarity with genes of Physarum polycephalum and Dictyostelium discoideum, which encode actin bundling proteins. Domain search analysis revealed the presence of essential conserved regions, i.e., an active actin binding site and 2 putative calcium binding EF-hands. Transfected amoeba cells demonstrated that AhABP is primarily localized in phagocytic cups, peripheral edges, pseudopods, and in cortical cytoplasm where actins are most abundant. Moreover, AhABP after the deletion of essential regions formed ellipsoidal inclusions within transfected cells. High-speed co-sedimentation assays revealed that AhABP directly interacted with actin in the presence of up to $10{\mu}M$ of calcium. Under the electron microscope, thick parallel bundles were formed by full length AhABP, in contrast to the thin actin bundles formed by constructs with deletion sites. In the light of these results, we conclude that AhABP is a novel actin bundling protein that is importantly associated with actin filaments in the cytoplasm.

Structural, Electrochemical, DNA Binding and Cleavage Properties of Nickel(II) Complex [Ni(H2biim)2(H2O)2]2+ of 2,2'-Biimidazole

  • Jayamani, Arumugam;Thamilarasan, Vijayan;Ganesan, Venketasan;Sengottuvelan, Nallathambi
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3695-3702
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    • 2013
  • A nickel(II) complex $[Ni(H_2biim)_2(H_2O)_2](ClO_4)_2{\cdot}H_2O$ (1) of biimidazole ligand has been synthesized and characterized (Where $H_2biim$ = 2,2'-biimidazole). The single crystal X-ray diffraction of the complex shows a dimeric structure with six coordinated psudo-octahedral geometry. The cyclic voltammograms of complex exhibited one quasireversible reduction wave ($E_{pc}=-0.61V$) and an irreversible oxidation wave ($E_{pa}=1.28V$) in DMF solution. The interaction of the complex with Calf-Thymus DNA (CT-DNA) has been investigated by absorption and fluorescence spectroscopy. The complex is an avid DNA binder with a binding constant value of $1.03{\times}10^5M^{-1}$. The results suggest that the nickel(II) complex bind to CT-DNA via intercalative mode and can quench the fluorescence intensity of EB bind to CT-DNA with $K_{app}$ value of $3.2{\times}10^5M^{-1}$. The complex also shown efficient oxidative cleavage of supercoiled pBR322 DNA in the presence of hydrogen peroxide as oxidizing agent. The DNA cleavage by complex in presence of quenchers; viz. DMSO, KI, $NaN_3$ and EDTA reveals that hydroxyl radical or singlet oxygen mechanism is involved. The complex showed invitro antimicrobial activity against four bacteria and two fungi. The antimicrobial activity was nearer to that of standard drugs and greater than that of the free ligand.

NEMO Support Scheme within PMIPv6 for Supporting Network Mobility in Harbor Area (항만 내 네트워크의 이동성 지원을 위한 PMIPv6망의 NEMO 지원 기법)

  • Min, Sang-Won;Han, Kyeong-Il;Jeong, Min-A;Lee, Seong Ro
    • The Journal of Korean Institute of Communications and Information Sciences
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    • v.39C no.8
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    • pp.674-680
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    • 2014
  • In this paper, as one of the fusion of IT and ocean technology, we propose a scheme to support NEMO in a harbor area network to provide network mobility for passengers, distribution as well as give a connection to an MN from an outside network. In a harbor area under PMIPv6 with NEMO support, a problem could be occurred due to overlapping HA/LMA tunnels and duplicate BCE fields. To Solve these problems, we consider the existing mobility option and add an optional field in the PBU message, which can notify the LMA and HA for either an initial connection or handover request. This procedure can reduce some overlapped steps of binding procedure. Hence, our proposed scheme could reduce an amount of delay and save the processing resource in the harbor area under PMIPv6 with NEMO support.

Inhibition Effects of Galla Chinenisis Extract on Adipocyte Differentiation in OP9 Cells (오미자 추출물의 지방세포 분화 억제 효과)

  • Park, Sun-Young;Hwang, Hong-Yeon;Seo, Eun-A;Kwon, Kang-Beom;Ryu, Do-Gon
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.26 no.4
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    • pp.455-461
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    • 2012
  • Obesity is associated with numerous diseases such as type 2 diabetes, hypertension and cancer. Inhibition of adipogenesis is a effectite strategy to anti-obesity. In this study, Galla Chinenisis extract (GCE) inhibited adipocyte differentiation in OP9 cells. There was no cytotoxicity when cells were treated with GCE in designated time intervals, unaffected by concentration. In this cell model, increases in fat storage were inhibited by 2 days treatment with various concentration of GCE, visualized by Oil red-O, BODIPY and DAPI staining. To understand the underlying mechanism at the molecular level, the effects of GCE were examined on the expression of the genes involved in adipogenesis by real-time PCR. In the progress of adipocyte differentiation with GCE-treated, the mRNA level of adipogenic genes such as peroxisome-proliferator-activated receptors gamma ($PPAR{\gamma}$), computer-assisted axial tomography/enhancer binding protein-alpha ($C/EBP{\alpha}$) were decreased. Also, GCE treatment inhibited increase of mRNA expression, which is adipogenic factor such as fatty acid synthase (FAS), hormone-sensitve lipase (HSL), lipoprotein lipase (LPL), and adipocyte-specific lipid binding protein (aP2). Therefore, the result of this study suggest that Galla Chinenisis extract can prevent adipocyte differentiation and GCE may have a great potential as a novel anti-adipogenic agent.

The Characterization of Anti-HER-2/neu Monoclonal Antibody using Different in vivo Imaging Techniques

  • Moon, Cheol;Kim, Eun Jung;Choi, Dan Bee;Kim, Byoung Soo;Kim, Sa Hyun;Choi, Tae Hyun
    • Biomedical Science Letters
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    • v.21 no.1
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    • pp.23-31
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    • 2015
  • Recently, specific antibodies have been used extensively to diagnose and treat various diseases. It is essential to assess the efficacy and specificity of antibodies, especially the in vivo environment. Anti-HER-2/neu mAb was evaluated as a possible transporting agent for radioimmunotherapy. The monoclonal antibody was successfully radio-labeled with $^{131}I$. In vitro binding assays were performed to confirm its targeting ability using another radio-iodine, $^{125}I$. Binding percentage of $^{125}I$ labeled anti-HER-2/neu mAb in HER-2/neu expressing CT-26 cells was found to be 4.5%, whereas the binding percentage of $^{125}I$ labeled anti-HER-2/neu mAb in wild-type CT-26 was only 0.45%. In vivo images were obtained and analyzed through $\gamma$-camera and an optical fluorescent modality, IVIS-200. $\gamma$-camera images showed that $^{131}I$ labeled anti-HER-2/neu mAb accumulated in HER-2/neu CT-26 tumors. Optical imaging based on near infrared fluorescence labeled anti-HER-2/neu mAb showed higher fluorescence intensities in HER-2/neu CT-26 tumors than in wild-type CT-26 tumors. Anti-HER-2/neu mAb was found to specifically bind to its receptor expressing tumor. Our study demonstrates that in vivo imaging technique is a useful method for the evaluation of an antibody's therapeutic and diagnostic potentials.

Testing of Cancer Chemopreventive Potential of Prunella vulgaris L. Aqua-acupuncture Solution Using Biochemical Markers of Carcinogenesis (발암과정 생화학적 표식자를 이용한 하고초 약침액의 암예방 활성 측정)

  • Park, Sin-Hwa;Cho, Kyoung-Hee;Shon, Yun-Hee;Lim, Jong-Kook;Nam, Kyung-Soo
    • Korean Journal of Pharmacognosy
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    • v.32 no.2 s.125
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    • pp.163-167
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    • 2001
  • Prunella vulgaris L. aqua-acupuncture solution (PVAS) was tested for cancer chemopreventive activity using chemoprevention-associated biochemical end points. The following effects were measured. : (a) inhibition of 7,12-dimethylbenz[a]anthracene (DMBA)-induced cytochrome P4501A1 activity (b) inhibition of $[^3H]B[a]P-DNA$ binding (c) inhibition of phorbol 12-myristate 13-acetate (TPA)-induced free radical formation in HL-60 cells (d) inhibition of polyamine metabolism. PVAS inhibited cytochrome P4501A1-mediated ethoxyresorufin-O-deethylase activity. The binding of $[^3H]B[a]P$ metabolites to DNA of NCTC-clone 1469 cells was inhibited significantly by PVAS. There is 22% inhibition of TPA-induced free radical formation in human leukemic cells with 5 mg/ml PVAS. Proliferation of Acanthamoeba castellanii was inhibited by PAVS at concentration of 30 mg/ml. PAVS positive in these assays may inhibit the carcinogenesis process and is considered very promising cancer-preventing agent because of its multiple activities.

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