• 제목/요약/키워드: glycoprotein D

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The Progression of SARS Coronavirus 2 (SARS-CoV2): Mutation in the Receptor Binding Domain of Spike Gene

  • Sinae Kim;Jong Ho Lee;Siyoung Lee;Saerok Shim;Tam T. Nguyen;Jihyeong Hwang;Heijun Kim;Yeo-Ok Choi;Jaewoo Hong;Suyoung Bae;Hyunjhung Jhun;Hokee Yum;Youngmin Lee;Edward D. Chan;Liping Yu;Tania Azam;Yong-Dae Kim;Su Cheong Yeom;Kwang Ha Yoo;Lin-Woo Kang;Kyeong-Cheol Shin;Soohyun Kim
    • IMMUNE NETWORK
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    • 제20권5호
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    • pp.41.1-41.11
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    • 2020
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense single-stranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor - angiotensin converting enzyme 2 (ACE2) - on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene. The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.

Biological Activity of Human Dimeric Hyperglycosylated Erythropoietin (dHGEPO) Fusion Proteins

  • Naidansuren, Purevjargal;Min, Kwan-Sik
    • Reproductive and Developmental Biology
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    • 제34권4호
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    • pp.289-297
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    • 2010
  • Erythropoietin (EPO) is a glycoprotein hormone secreted from primarily cells of the peritubular capillary endothelium of the kidney, and is responsible for the regulation of red blood cell production. We constructed and expressed dimeric cDNAs in Chinease hamster ovary (CHO) cells encoding a fusion protein consisting of 2 complete human EPO domains linked by a 2-amino acid linker (Ile-Asp). We described the activity of dimeric hyperglycosylated EPO (dHGEPO) mutants containing additional oligosaccharide chains and characterized the function of glycosylation. No dimeric proteins with mutation at the $105^{th}$ amino acid were found in the cell medium. Growth and differentiation of the human EPO-dependent leukemiae cell line (F36E) were used to measure cytokine dependency and in vitro bioactivity of dHGEPO proteins. MIT assay at 24 h increased due to the survival of F36E cells. The dHGEPO protein migrated as a broad band with an average molecular mass of 75 kDa. The mutant, dHGEPO, was slightly higher than the wild-type (WT) dimeri-EPO band. Enzymatic N-deglycosylation resulted in the formation of a narrow band with a molecular mass twice of that of of monomeric EPO digested with an N-glycosylation enzyme. Hematocrit values were remarkably increased in all treatment groups. Pharmacokinetic analysis was also affected when 2.5 IU of dHGEPO were intravenously injected into the tails of the mice. The biological activity and half-life of dHGEPO mutants were enhanced as compared to the corresponding items associated the WT dimeric EPO. These results suggest that recombinant dHGEPO may be attractive biological and therapeutic targets.

설치류에서 알파 Herpes 바이러스의 신경친화성과 침습 (Neurotropism and Invasiveness of $\alpha-Herpes$ Virus in the Rodent)

  • 김진상;이성준
    • The Journal of Korean Physical Therapy
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    • 제9권1호
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    • pp.59-70
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    • 1997
  • The ability of neurotropic alpha herpesviruses to replicate within synaptically linked neurons has made these pathogens valuable tools for transneuronal analysis. Recent studies suggest that unique gene products expressed by genetically engineered strains of virus may permit the use of multiple strains in complex tracing paradigms. In the present study we have examined the invasiveness of two genetically engineered strains of the swine pathogen known as pseudorabies virus(PRV). The two strains were isogenic with the attenuated Bartha strain of PRV; in one strain a lacZ reporter gene was inserted into the gC locus (PRV-BaBlu; $4.75\times10^8pfu/ml$) contrained a PRV envelope glycoprotein gene that was absent in PRV-BaBlu. Simultaneous or temporally separated sequential injection of $4\mu\ell$ of each strain into the ventral wall of the stomach produced a predictale course of retrograde synaptic infection. The results were as follows: 1. PRV-BaBlu and PRV-D infected the dorsal motor nucleus of vagus nerve(DMV) and paraventricular nucleus(PVN). 2. Invasion and replication of PRV-D occured at a faster rate than the parental strain or PRV-BaBlu. 3. PRV-D was much more virulent than PRV-BaBlu or the parental strain. 4. Co-injection of PRV-D and PRV-BaBlu produced an infection that was more virulent than that produced by the parental strain (PRV-Bartha), 5. Neurons in DMV were permissive to co-infection with PRV-D and PRV-BaBlu when they were injected simultaneously into the same site. 6. Replication of PRV-BaBlu was compromised by prior infection of the same circuit with PRV-D. 7. Prior infection of neurons with PRV-D maked them resistant to infection with PRV-BaBlu.

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Cell-Surface Loss of Constitutive Activating and Inactivating Mutants of Eel Luteinizing Hormone Receptors

  • Byambaragchaa, Munkhzaya;Choi, Seung-Hee;Kim, Dong-Wan;Min, Kwan-Sik
    • 한국발생생물학회지:발생과생식
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    • 제25권4호
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    • pp.225-234
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    • 2021
  • The present study aimed to investigate the mechanism of cell surface receptor loss by two constitutively activating mutants (designated L469R, and D590Y) and two inactivating mutants (D417N and Y558F) of the luteinizing hormone receptor (LHR) in the Japanese eel Anguilla japonica, known to naturally occur in human LHR transmembrane domains. We investigated cell surface receptor loss using an enzyme-linked immunosorbent assay in HEK 293 cells. The expression level of wild-type eel LHR was considered to be 100%, and the expression levels of L469R and D417N were 97% and 101%, respectively, whereas the expression levels of D590Y and Y558F slightly increased to approximately 110% and 106%, respectively. The constitutively activating mutants L469R and D590Y exhibited a decrease in cell surface loss in a manner similar to that of wild-type eel LHR. The rates of loss of cell surface agonist-receptor complexes were observed to be very rapid (2.6-6.2 min) in both the wild-type eel LHR and activating mutants. However, cell surface receptor loss in the cells expressing inactivating mutants D417N and Y558F was slightly observed in the cells expressing inactivating mutants D417N and Y558F, despite treatment with a high concentration of agonist. These results provide important information on LHR function in fish and the regulation of mutations of highly conserved amino acids in glycoprotein hormone receptors.

FERTILITY STUDY (SEGMENT 1) OF RECOMBINANT GRANULOCYTE-MACROPHAGE COLONY STIMULATING FACTORS (LBD-005) IN RATS

  • Kim, Sung-Hoon;Chung, Moon-Koo;Roh, Jung-Koo
    • Toxicological Research
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    • 제8권1호
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    • pp.29-40
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    • 1992
  • The recombinant glycoprotein, LBD-005 (Lucky R & D Center, Biotechnology)stimulates the growth of stem cells and activates the development of the hematopoietic cell in a similar manner to the naturally occurring GM-CSF. This test was conducted to investigate if LBD-005 had any effect on fertility in male and female rats. Sprague-Dawley rats(88 of each sex) bred at KRICT, were dosed subcutaneously, at a volume of 2ml/kg body weight with LBD-005 at 0, 250, 500 or 1, 000mg/kg body weight. The male rats were dosed, from 6 weeks of asge, for 60 days prior to mating.

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저근백피의 Virus-Cell Fusion 저해활성 성분 (Virus-Cell Fusion Inhibitory Compounds from Ailanthus altissima Swingle)

  • 장영수;문영희;우은란
    • 생약학회지
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    • 제34권1호통권132호
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    • pp.28-32
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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. Repeated column chromatoghaphy of the methylene chloride fraction of A. altissima afforded compounds 1$({\beta}-sitosterol-3-O-{\beta}-D-glucoside)$, 2(tetramethoxycoumarin), and 3(ocotillone). Virus-cell fusion inhibitory activity of compound 3(ocotillone) was $70.76{\pm}4.09%$ at the concentration of $100\;{\mu}g/ml$.

Purification and In Vitro Translation of Penicillium verruculosum Cellulase mRNA

  • Kim, Jeong-Ho;Chung, Ki-Chul;Kang, Hyun-Sam;Lee, Young-Kyu
    • Journal of Microbiology and Biotechnology
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    • 제1권4호
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    • pp.232-239
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    • 1991
  • Caboxymethyl cellulase (CMCase) I was purified from the induced culture filtrate of Penicllium verruculosum F-3 by ammonium sulfate precipitation, DEAE-Sephadex A-50 chromatography and Bio-gel P-150 filtration. The purified enzyme was assumed to be a glycoprotein consisting of 8.5% carbohydrate and having a molecular weight of 70.000 in SDS-polycrylamide gel electrophoresis (SDS-PAGE). The purified enzyme-specific anti-CMCase I IgG was obtained by rabbit immunization and protein A-sepharose CL-4B chromatography. The fungal poly($A^+$) RNA was isolated from the total RNA of the mycelium grown under cellulase induction conditions by oligo(dT)-cellulosse chromatography. The translation products in vitro were prepared by translating the isolated poly ($A^+$) RNA in rabbit reticulocyte lysate and analyzed by SDS-PAGE and fluorography. Of the translation products, CMCase I was identified by the immunoprecipitation against anti-CMCase I IgG.

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Investigation of Binding Modes of the Verapamil and Curcumin into Human P-glycoprotein (P-gp)

  • Gadhe, Changdev G.;Cho, Seung Joo
    • 통합자연과학논문집
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    • 제6권4호
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    • pp.205-210
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    • 2013
  • Human P-gp is a protein responsible for the multidrug resistance (MDR) and causes failure of cancer chemotherapy. Till date no X-ray crystal structure is reported for this membrane protein, which hampers active research in the field. We performed homology modeling to develop three dimensional (3D) model of P-gp, and docking studies of the verapamil and curcumin have been performed to gain insight into the interaction mechanism between inhibitors and P-gp. It was identified that the inhibitors docked into the upper part of P-gp and interacted through the hydrophobic interactions.

Chemical Dissection of Zebrafish Egg Envelop, the Chorion

  • Hwang, C. N.;H. J. Kang;Kim, C.;D. S. Na;S. K. Chae;B. K. Joo;Lee, J. W.;Lee, S. H.
    • 한국동물번식학회:학술대회논문집
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    • 한국동물번식학회 2004년도 춘계학술발표대회
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    • pp.283-283
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    • 2004
  • The eggs of most animal species are surrounded by an extracellular matrix known as chorion, egg envelope, egg coat, or zona pellucida. Development of fish embryo usually takes several days in an aquatic environment. During embryonic development, the chorion must protect embryo from physical damage and microbial infection in the exposed aquatic environment. (omitted)

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Effects of heat stress on growth performance, selected physiological and immunological parameters, caecal microflora, and meat quality in two broiler strains

  • Awad, Elmutaz Atta;Najaa, Muhamad;Zulaikha, Zainool Abidin;Zulkifli, Idrus;Soleimani, Abdoreza Farjam
    • Asian-Australasian Journal of Animal Sciences
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    • 제33권5호
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    • pp.778-787
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    • 2020
  • Objective: This study was conducted to investigate the effects of normal and heat stress environments on growth performance and, selected physiological and immunological parameters, caecal microflora and meat quality in Cobb 500 and Ross 308 broilers. Methods: One-hundred-and-twenty male broiler chicks from each strain (one-day-old) were randomly assigned in groups of 10 to 24 battery cages. Ambient temperature on day (d) 1 was set at 32℃ and gradually reduced to 23℃ on d 21. From d 22 to 35, equal numbers of birds from each strain were exposed to a temperature of either 23℃ throughout (normal) or 34℃ for 6 h (heat stress). Results: From d 1 to 21, strain had no effect (p>0.05) on feed intake (FI), body weight gain (BWG), or the feed conversion ratio (FCR). Except for creatine kinase, no strain×temperature interactions were observed for all the parameters measured. Regardless of strain, heat exposure significantly (p<0.05) reduced FI and BWG (d 22 to 35 and 1 to 35), immunoglobulin Y (IgY) and IgM, while increased FCR (d 22 to 35 and 1 to 35) and serum levels of glucose and acute phase proteins (APPs). Regardless of temperature, the Ross 308 birds had significantly (p<0.05) lower IgA and higher finisher and overall BWG compared to Cobb 500. Conclusion: The present study suggests that the detrimental effects of heat stress are consistent across commercial broiler strains because there were no significant strain×temperature interactions for growth performance, serum APPs and immunoglobulin responses, meat quality, and ceacal microflora population.