• Title/Summary/Keyword: gp41

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Isolation of the Gene for HIV-1 gp41 Interacting Protein (HIV gp41의 세포내 부분과 상호작용하는 단백질 유전자의 분리)

  • Kim, Eun-Mi;Kim, Jung-Woo
    • The Journal of Natural Sciences
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    • v.10 no.1
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    • pp.27-32
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    • 1998
  • To find the interacting protein with the cytoplasmic domain of HIV-1 gp41, the yeast two hybrid system was used for the expression cloning. Among the $1.4 \times 10^6 colonies, 20 colonies were selected as the final candidate for the interacting protein gene. The nucleotide sequencing revealed three kinds of protein, acidic ribosomal protein P0, beta tubulin, alpha catenin. These proteins interacted with the gp41 specifically in yeast system.

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Characterization and Epitope Mapping of KI-41, a Murine Monoclonal Antibody Specific for the gp41 Envelope Protein of the Human Immunodeficiency Virus-1

  • Shin, Song-Yub;Park, Jung-Hyun;Jang, So-Youn;Lee, Myung-Kyu;Hahm, Kyung-Soo
    • BMB Reports
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    • v.31 no.1
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    • pp.58-63
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    • 1998
  • In this study, a mouse monoclonal antibody (mAb) against gp41(584-618), the immunodominant epitope protein, was generated. For this purpose, BALB/c mice were immunized with double branched multiple antigenic peptides derived from the HIV-1 gp41(584-618) sequence, and antibody-secreting hybridoma were produced by fusion of mice splenocytes with SP2/0 myeloma cells. One clone producing an antigen specific mAb, termed KI-41(isotype IgG1) was identified, whose specific reactivity against gp41(584-618) could be confirmed by ELISA and Western blot analysis. Epitope mapping revealed the recognition site of the mAb KI-41 to be located around the sequence RILAVERYLKDQQLLG, which comprises the N-terminal region within the immunized gp41(584-618) peptied. Since this mAb recognizes this specific epitope within the HIV-1 gp41 without any cross-reactivity to other immunodominant regions in the HIV-2 gp35, KI-41 will provide some alternative possibilities in further applications such as the development of indirect or competitive ELISA for specific antibody detection in HIV-1 infection or for other basic researches regarding the role and function of HIV-1 gp41.

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Membrane interaction of the coiled-coil motif of HIV gp41 and its implication in the membrane fusion process

  • Jin, Bong-Suk;Yu, Yeon-Gyu
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.58-58
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    • 2003
  • The envelope glycoprotein of HIV, gp41, mediates the membrane fusion with human cells. The extracellular domain of gp41 has two helical regions. The N-terminus helical region (N-helix) forms trimeric coiled coil, interacts with the C-terminus helical region (C-helix) of gp41 to form a stable helical bundle structure. In this study, we have shown that the N-helix of gp41 has membrane interacting and disrupting abilities. It was localized into the interface of the lipidic phase and head group of the membrane. In contrast, the N-helix region with membrane fusion defective mutations could not bind to membrane. In addition, the N-helix bound on the membrane was released from the membrane by the C-helix, and the complex of the N- and C-helix did not interact with membrane. These results suggested that the membrane binding ability of the N-helix is necessary for the fusion activity of gp41, and such property is possibly controlled by the C-helm.

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Expression and Characterization of Human Immunodeficiency Virus-1 Oligomerized gp140 Protein in Mammalian Cells (포유동물 세포에서 Human Immunodeficiency Virus-1의 Oligomeric gp140 단백의 발현 및 특성)

  • Kim, Eun-Ok;Kim, Eun;Kim, Hyun-Soo;Shin, Kwang-Soon;Kim, Chul-Joong
    • Korean Journal of Veterinary Research
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    • v.42 no.1
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    • pp.55-64
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    • 2002
  • The envelope glycoprotein of HIV-1 forms an oligomeric complex resulting in playing a role to induce neutralizing antibody and cell-mediate immune responses. The oligomer exists as a trimer of gp120-gp41 heterodimer which mediates HIV-1 attachment and fusion. We made a cDNA clone of gp140 consisting of gp120 and ectodomain of gp41 from the primary African isolate. To express the oligomeric gp140 in mammalian cells, we adopted the Semliki Forest virus (SFV) based expression system. The oligomeric gp140 in the secretory form was expressed and purified from the cell culture supernatant and characterized. The antibody inducing activity of the purified gp140 was also examined in mice inoculation.

Structural Analysis of the Ectodomain of HIV Gp41 and Implication on the Gp41 Assisted Membrane Fusion

  • Ryu, Jae-Ryen;Lee, Jung;Suh, Mu-Jin;Yu, Yeong-Sook;Yu, Yeon-Gyu
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.33-33
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    • 1996
  • An ectodomain of gp41, the transmembrane fusion protein of HIV, without the fusion peptide region was expressed using pET system in E. coli. The expressed protein gp41core, was isolated as inclusion body and was purified by ion-exchange chromatography after solubilized in 6M urea. The purified denatured protein was renaturated and the folded domain of gp41core was identified by the presence of the proteolysis resistence domain and a high content of ${\alpha}$-helical secondary structure. (omitted)

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Overexpression and Purification of p24 and gp41 Proteins of Human Immunodeficiency Virus Type 1 in E. coli (대장균에서 인간면역결핍 바이러스 1형의 gag p24 및 env gp41 단백질의 과발현 및 정제)

  • Kim, Chae-Young;Shin, Soon-Cheon;Lee, Sung-Hee;Kim, Won-Bae;Kim, Byong-Moon
    • The Journal of Korean Society of Virology
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    • v.28 no.1
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    • pp.21-30
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    • 1998
  • Synthetic genes encoding the gag p24 and the part of the envelope protein gp41 of the human immunodeficiency virus (HIV-1) were cloned and overexpressed as fusion proteins in Escherichia coli, using an expression vector carrying T7 promoter and the poly-histidine leader, sequence. The overexpressed p24 fusion protein was purified by centrifugation, Ni-affinity chromatography and CM-sepharose chromatography. The overexpressed gp41 fusion protein was purified by centrifugation, $C_4$ chromatography and DEAE-sepharose chromatography. The purified fusion proteins showed a high level of purity and immunoreactivity in SDS-polyacrylamide gel electrophoresis and western blot analysis. These results suggest that this prokaryotic - expression purification method is suitable for obtaining a large amount of the viral antigen which may be useful for screening of antibodies to HIV-1 in human blood samples.

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Implementation of parallel blocked LU decomposition program for utilizing cache memory on GP-GPUs (GP-GPU의 캐시메모리를 활용하기 위한 병렬 블록 LU 분해 프로그램의 구현)

  • Kim, Youngtae;Kim, Doo-Han;Yu, Myoung-Han
    • Journal of Internet Computing and Services
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    • v.14 no.6
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    • pp.41-47
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    • 2013
  • GP-GPUs are general purposed GPUs for numerical computation based on multiple threads which are originally for graphic processing. GP-GPUs provide cache memory in a form of shared memory which user programs can access directly, unlikely typical cache memory. In this research, we implemented the parallel block LU decomposition program to utilize cache memory in GP-GPUs. The parallel blocked LU decomposition program designed with Nvidia CUDA C run 7~8 times faster than nun-blocked LU decomposition program in the same GP-GPU computation environment.

Expression and Characterization of the Human Immunodeficiency Virus Type 1 Mutant Envelope Glycoproteins in Mammalian Cells (진핵세포에서 HSV-1 Envelope 변이 단백질의 발현 및 발현 단백질의 특성 연구)

  • Ryu, Ji-Yoon;Park, Jin-Seu
    • The Journal of Korean Society of Virology
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    • v.29 no.3
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    • pp.183-193
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    • 1999
  • Human immunodeficiency virus type 1 (HIV-1) envelope glycoprotein is synthesized as a 160 KDa precursor, gp160, that is cleaved by a cellular protease to form the gp120 and gp41 subunits. Mammalian expression vectors were designed that are capable of efficient expression of various mutant envelope glycoproteins derived from a molecular clone of HIV-1. To construct these vectors, one type of mutation was made at the gp120-gp41 cleavage site by oligonucleotide-directed mutagenesis. And another mutation was made to change amino acids in the membrane spanning region of HIV-1 gp41 important for membrane anchorage. Next, these two mutations were combined to generate a vector to have double mutations in cleavage site and membrane-spanning region. These mutants were transiently expressed in mammalian cells. The effect of these mutations on envelope glycoprotein synthesis, proteolytic processing and secretion was determined. In addition, cell surface expression and ability of the glycoprotein to induce syncytium formation were examined. This study provides a mammalian expression system that is capable of efficient expression and secretion of soluble gp160.

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Combination of Curcumin and Paclitaxel-loaded Solid Lipid Nanoparticles to Overcome Multidrug Resistance

  • Li, Rihua;Xu, Wenting;Eun, Jae-Soon;Lee, Mi-Kyung
    • Journal of Pharmaceutical Investigation
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    • v.41 no.6
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    • pp.381-386
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    • 2011
  • Multi-drug resistance (MDR) has been known as a major hurdle in cancer chemotherapy. One of the most clinically significant causes of MDR was the efflux of anticancer agents mediated by p-glycoprotein (p-gp) over-expressed in MDR cancer cells. To overcome MDR, there have been several strategies such as co-administration with p-gp inhibitors and encapsulation of anticancer drugs into drug delivery systems. In the present study, curcumin was evaluated for its potential as p-gp inhibitor and MDR reversal activity when combined with paclitaxel incorporated into lipid nanoparticles (PTX/LN). Western blot assay showed curcumin did not modulate the level of p-gp expression in MCF-7/ADR which is a MDR variant of human breast cancer cell line, MCF-7, and over-expresses p-gp. However, curcumin inhibited p-gp-mediated efflux of calcein in a dose-dependent manner even though it showed lower activity compared to verapamil, a well-known p-gp inhibitor. Incorporation of paclitaxel into lipid nanoparticles partially recovered the anticancer activity of paclitaxel in MCF-7/ADR. The combined use of curcumin and PTX/LN exhibited further full reversal of MDR, suggesting susceptibility of PTX/LN to the efflux system. In conclusion, combined approach of using p-gp inhibitors and incorporation of the anticancer agents into nano-delivery systems would be an efficient strategy to overcome MDR.

GP-GPU based Parallelization for Urban Terrain Atmospheric Model CFD_NIMR (도시기상모델 CFD_NIMR의 GP-GPU 실행을 위한 병렬 프로그램의 구현)

  • Kim, Youngtae;Park, Hyeja;Choi, Young-Jeen
    • Journal of Internet Computing and Services
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    • v.15 no.2
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    • pp.41-47
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    • 2014
  • In this paper, we implemented a CUDA Fortran parallel program to run the CFD_NIMR model on GP-GPU's, which simulates air diffusion on urban terrains. A GP-GPU is graphic processing unit in the form of a PCI card, and a general calculation accelerator to perform a large amount of high speed calculations with low cost and electric power. The GP-GPU gives performance enhancement of speed by 15 times to compare the Nvidia Tesla C1060 GPU with Intel XEON 2.0 GHz CPU. In addition, the program on a GP-GPU shows efficient performance compared to an MPI parallel program on multiple CPU's. It is expected that a proposed programming method on the GP-GPU parallel program can be used for numerical models with a similar structure.