• Title/Summary/Keyword: envelope protein

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Computational Prediction of Solvation Free Energies of Amino Acids with Genetic Algorithm

  • Park, Jung-Hum;Lee, Jin-Won;Park, Hwang-Seo
    • Bulletin of the Korean Chemical Society
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    • v.31 no.5
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    • pp.1247-1251
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    • 2010
  • We propose an improved solvent contact model to estimate the solvation free energies of amino acids from individual atomic contributions. The modification of the solvation model involves the optimization of three kinds of parameters in the solvation free energy function: atomic fragmental volume, maximum atomic occupancy, and atomic solvation parameters. All of these atomic parameters for 17 atom types are developed by the operation of a standard genetic algorithm in such a way to minimize the difference between experimental and calculated solvation free energies. The present solvation model is able to predict the experimental solvation free energies of amino acids with the squared correlation coefficients of 0.94 and 0.93 for the parameterization with Gaussian and screened Coulomb potential as the envelope functions, respectively. This result indicates that the improved solvent contact model with the newly developed atomic parameters would be a useful tool for the estimation of the molecular solvation free energy of a protein in aqueous solution.

Identifying immunodominant multi-epitopes from the envelope glycoprotein of the Lassa mammarenavirus as vaccine candidate for Lassa fever

  • Adekunle Babajide Rowaiye;Ezinne Janefrances Nwonu;Titilayo Mercy Asala;Amoge Chidinma Ogu;Doofan Bur;Chimaobi Chukwu;Angus Nnamdi Oli;Tarimoboere Agbalalah
    • Clinical and Experimental Vaccine Research
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    • v.11 no.3
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    • pp.249-263
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    • 2022
  • Purpose: Lassa fever is a zoonotic acute viral hemorrhagic disease caused by Lassa virus (LASV). There is currently no licensed vaccine for the prevention of the disease. This study is aimed at discovering immunodominant epitopes from the envelope glycoprotein of the Lassa mammarenavirus and designing of a multi-epitope vaccine candidate (VC). Materials and Methods: The amino acid sequences of the envelope glycoprotein of 26 strains of LASV from five countries were selected. After evaluation for antigenicity, immunogenicity, allergenicity, and toxicity, immunodominant CD8, CD4, and linear B lymphocytes were also selected. The selected epitopes were modelled and a molecular docking with the appropriate major histocompatibility complex (MHC) proteins was performed. Using an adjuvant and linkers, a multi-epitope VC was designed. The VC was evaluated for its physicochemical and immunological properties and structurally refined, validated, and mutated (disulphide engineering). The complex formed by the VC and the toll-like receptor-4 receptor was subjected to molecular dynamic simulation (MDS) followed by in silico cloning in a plasmid vector. Results: A VC with 203 sequences, 22.13 kDa weight, isoelectric point of 9.85 (basic), instability index value of 27.62, aliphatic index of 68.87, and GRAVY value of -0.455 (hydrophilic) emerged. The VC is predicted to be non-allergenic with antigenicity, MHC I immunogenicity, and solubility upon overexpression values of 0.81, 2.04, and 0.86 respectively. The VC also has an estimated half-life greater than 10 hours in Escherichia coli and showed stability in all the parameters of MDS. Conclusion: The VC shows good promise in the prevention of Lassa fever but further tests are required to validate its safety and efficacy.

Construction of a Fluorescently Labeled Infectious R Peptide-Less Moloney MLV Molecular Clone for Analysis of Syncytium (합포체 분석을 위해 R 펩타이드가 결여된 형광 표지 Moloney 마우스레트로바이러스 Molecular Clone 제조)

  • Lee, Yong-Jin;Park, Jin-Woo;Lee, Kyu-Jun;Bae, Eun-Hye;Park, Sung-Han;Lim, Ji-Hyun;Kim, Sae-Ro-Mi;Jung, Yong-Tae
    • Korean Journal of Microbiology
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    • v.45 no.3
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    • pp.246-250
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    • 2009
  • Retroviruses enter host cells by membrane fusion between the viral Env proteins on the virus membrane and a virus receptor on the cellular membrane. The envelope protein of the ecotropic Moloney murine leukemia virus is synthesized as a gp85 precursor and is proteolytically cleaved into an extracellular surface unit (SU) and the transmembrane protein (TM). The cytoplasmic tail (16 amino acid; R peptide) of the TM protein is further cleaved by the viral protease during virion maturation. Unlike the wild type Env protrin bearing the R peptide, R peptide-truncated Envelope induces syncytia in susceptible cells. To understand the mechanism of R peptidetruncated Env in syncytium formation, R peptide-truncated Env expressing full-length molecular clone containing EGFP in PRR (proline rich region) of Env was constructed. This molecular clone induced syncytia in transfected NIH3T3 cells, fluorescence was detected in the cytoplasm and at the plasma membrane, while the nuclei did not stain and appeared black by fluorescence microscopy. Interestingly, virions with truncated envelope produced from transfected NIH3T3 cells induced syncytia in NIH3T3 cells, but fluorescence was not detected in the same infected cells. It is believed that cell-free viruses direct the fusion of neighboring cells without infection. Our data suggests that use of EGFP-tagged envelope for monitoring syncytium is a sensitive and convenient method. We also found that virion incorporated the R peptide-truncated Env is able to induce the formation of syncytia by fusion from without.

Dose absorption of Omicron variant SARS-CoV-2 by electron radiation: Using Geant4-DNA toolkit

  • Mehrdad Jalili Torkamani;Chiman Karami;Pooneh Sayyah-Koohi;Farhood Ziaie;Seyyedsina Moosavi;Farhad Zolfagharpour
    • Nuclear Engineering and Technology
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    • v.56 no.6
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    • pp.2421-2427
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    • 2024
  • In this research, the Omicron variant of the SARS-CoV-2 virus was simulated and exposed to electron radiation with up to 20 keV energy. Absorbed energy was measured for spike protein, nucleocapsid protein, and envelope of the virus. Simulations were performed by Geant4-DNA in a water environment at temperature of 20 ℃ and pressure of 1 atm. Since the viral RNA is kept inside the nucleocapsid protein, damage to this area could destroy the viral RNA strand and create an inactive virus. Our findings showed that electron beams with an energy of 2.5 keV could cause a maximum absorption dose and consequently maximum damage to the nucleocapsid and effectively be used for inactivation virus.

N-Terminal Amino Acid Sequences of Receptor-Like Proteins that Bind to preS1 of HBV in HepG2 Cells

  • Lee, Dong-Gun;Liu, Ming-Zhu;Kim, Kil-Lyong;Hahm, Kyung-Soo
    • BMB Reports
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    • v.29 no.2
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    • pp.180-182
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    • 1996
  • One of the essential functions of virus surface proteins is the recognition of specific receptors on target cell membranes, and cellular receptors play an important role in viral pathogenesis. But the earliest steps of hepatitis B virus (HBV) infection, such as hepatocyte receptor interaction with the virus, are poorly understood. Previous work has suggested an important role of the preS1 region of HBV envelope protein in mediating viral binding to hepatocytes. Although hepatitis B virus (HBV) infection appears to be initiated by specific binding of virions to cell membrane structures via one or potentially several viral surface proteins, data showing the identification or isolation of the HBV receptor (s) are not yet available. The receptor-like proteins on the plasma membrane surface of HepG2 cells that bind to PreS1 were separated and identified using affinity chromatography, and the amino-terminal amino acid sequences of the receptor-like proteins were determined.

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Immunohistochemistry for detection of Aujeszky's disease virus antigens: Protein A-gold labeling of ultrathin sections for electron microscopy (오제스키병 바이러스 항원검출을 위한 면역조직화학적 연구 : 전자현미경적 관찰을 위한 초박절편내 protein A-gold labeling)

  • Kim, Soon-bok
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.541-548
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    • 1989
  • The present study was carried out to determine viral antigens and its morphogenesis in the ultrathin frozen and araldite sections of cell cultures infected with ADV by protein A-gold labeling. ADV antigens were labeled with 10nm gold probes, and electron-dense gold particles were mainly present on viral nucleocapsids and viral envelopes. Immunogold labeling in the ultracryosections showed a very low degree of interaction with tissue structures. Immunogold labeling in the ultrathin cryosections can be useful tool for the detection of ADV antigens, and the technique also may provide its great potential for immunocytochemical studies on various virus-host cell Interactions.

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Virus-cell fusion inhibitory compounds from Ailanthus altissima Swingle

  • Lee, Hyang-Hee;Chang, Young-Su;Moon, Young-Hee;Woo, Eun-Rhan
    • Proceedings of the PSK Conference
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    • 2003.04a
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    • pp.264.1-264.1
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    • 2003
  • In order to search for the anti-HIV agents from natural products, Eighty MeOH extracts of medicinal plants were applied to a syncytia formation inhibition assay which is based on the interaction between the HIV-1 envelope glycoprotein gp120/gp41 and the cellular membrane protein CD4 of T lymphocytes. Among them, Ailanthus altissima showed a potent virus-cell fusion inhibitory activity. (omitted)

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Emerging Roles of CTD Phosphatases (CTD 탈 인산화 효소의 기능과 역할)

  • Kim, Youngjun
    • Journal of Life Science
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    • v.27 no.3
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    • pp.370-381
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    • 2017
  • Protein dephosphorylation is important for cellular regulation, which is catalyzed by protein phosphatases. Among protein phosphatases, carboxy-terminal domain (CTD) phosphatases are recently emerging and new functional roles of them have been revealed. There are 7 CTD phosphatases in human genome, which are composed of CTD phosphatase 1 (CTDP1), CTD small phosphatase 1 (CTDSP1), CTD small phosphatase 2 (CTDSP2), CTD small phosphatase-like (CTDSPL), CTD small phosphatase-like 2 (CTDSPL2), CTD nuclear envelope phosphatase (CTDNEP1), and ubiquitin-like domain containing CTD phosphatase 1 (UBLCP1). CTDP1 dephosphorylates the second phosphor-serine of CTD of RNA polymerase II (RNAPII), while CTDSP1, STDSP2, and CTDSPL dephosphorylate the fifth phosphor-serine of CTD of RNAPII. In addition, CTDSP1 dephosphorylates new substrates such as mothers against decapentaplegic homologs (SMADs), cell division cycle-associated protein 3 (CDCA3), Twist1, tumor-suppressor protein promyelocytic leukemia (PML), and c-Myc. CTDP1 is related to RNA polymerase II complex recycling, mitosis regulation and cancer cell growth. CTDSP1, CTDSP2 and CTDSPL are related to transcription factor recruitment, tumor suppressor function and stem cell differentiation. CTDNEP1 dephosphorylates LIPIN1 and is related to neural tube formation and nuclear envelope formation. CTDSPL2 is related to hematopoietic stem cell differentiation. UBLCP1 dephosphorylates 26S proteasome and is related to nuclear proteasome regulation. In conclusion, noble roles of CTD phosphatases are emerging through recent researches and this review is intended to summarize emerging roles of CTD phosphatases.

Finding and Characterization of Viral Nonstructural Small Protein in Prospect Hill Virus Infected Cell

  • Nam, Ki-Yean;Chung, Dong-Hoon;Choi, Je-Won;Lee, Yun-Seong;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.29 no.4
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    • pp.221-233
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    • 1999
  • Prospect Hill Virus (PHV) is the well known serotype of hantavirus, a newly established genus in family Bunyaviridae. Extensive studies have upheld the original view of PHV genetics with three genes such as nucleocapsid (N) protein, envelope proteins (G1, G2) and RNA dependent RNA polymerase. In this study, we report the existence of additional gene that is encoded in an overlapping reading frame of the N protein gene within S genome segment of PHV. This gene is expected to encode a nonstructural small (NSs) protein and it seems to be only found in PHV infected cell. The presence and synthesis of NSs protein could be demonstrated in the cell infected with PHV using anti-peptide sera specific to the predicted amino acid sequence deduced from the second open reading frame. Ribosomal synthesis of this protein appears to occur at AUG codon at the 83rd base of S genome segment, downstream of N protein initiation codon. This protein is small in size (10.4 KDa) and highly basic in nature. The expression strategy of NSs protein appears that a signal mRNA is used to translate both N and NSs protein in PHV infected cell. 10 KDa protein in virus infected cell lysates can bind to mimic dsRNA. This fact strongly suggests that NSs protein may be involved in virus replication on late phase of viral life cycle.

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Defining the N-Linked Glycosylation Site of Hantaan Virus Envelope Glycoproteins Essential for Cell Fusion

  • Zheng, Feng;Ma, Lixian;Shao, Lihua;Wang, Gang;Chen, Fengzhe;Zhang, Ying;Yang, Song
    • Journal of Microbiology
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    • v.45 no.1
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    • pp.41-47
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    • 2007
  • The Hantaan virus (HTNV) is an enveloped virus that is capable of inducing low pH-dependent cell fusion. We molecularly cloned the viral glycoprotein (GP) and nucleocapsid (NP) cDNA of HTNV and expressed them in Vero E6 cells under the control of a CMV promoter. The viral gene expression was assessed using an indirect immunofluorescence assay and immunoprecipitation. The transfected Vero E6 cells expressing GPs, but not those expressing NP, fused and formed a syncytium following exposure to a low pH. Monoclonal antibodies (MAbs) against envelope GPs inhibited cell fusion, whereas MAbs against NP did not. We also investigated the N-linked glycosylation of HTNV GPs and its role in cell fusion. The envelope GPs of HTNV are modified by N-linked glycosylation at five sites: four sites on G1 (N134, N235, N347, and N399) and one site on G2 (N928). Site-directed mutagenesis was used to construct eight GP gene mutants, including five single N-glycosylation site mutants and three double-site mutants, which were then expressed in Vero E6 cells. The oligosaccharide chain on residue N928 of G2 was found to be crucial for cell fusion after exposure to a low pH. These results suggest that G2 is likely to be the fusion protein of HTNV.