• Title/Summary/Keyword: M protein

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THE INFLUENCE OF DIETARY PROTEIN AND ENERGY LEVELS ON EGG QUALITY IN STARCROSS LAYERS

  • Uddin, M. Salah;Tareque, A.M.M.;Howlider, M.A.R.;Khan, M. Jasimuddin
    • Asian-Australasian Journal of Animal Sciences
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    • v.4 no.4
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    • pp.399-405
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    • 1991
  • The interaction of 4 dietary crude protein (13, 16, 19 or 22%) and 4 metabolizable energy (2600, 2800, 3000 or 3100 kcal ME/kg) levels on egg quality performances of Starcross layers were assessed between 245 and 275 days of age. The egg weight increased significantly with the increasing dietary protein and energy levels. But egg shape index, albumen index, yolk index, yolk dry matter, yolk protein, yolk fat, albumen protein and shell tickness were similar at all dietary protein and/or energy levels. The egg specific gravity and albumen weight increased but the yolk, weight, Haugh unit and albumen drymatter decreased with the increase of dietary protein levels and showed irregular trend with energy levels. The albumen dry matter and egg shell weight, however, were not affected by energy and protein levels. Simultaneous increase of protein and energy increased specific gravity, albumen index and shell thickness at a greater rate than that increased by the increase of protein or energy alone.

Zeolite-Mediated Cation Exchange Enhances the Stability of mRNA during Cell-Free Protein Synthesis

  • Kim, You-Eil;Kim, Dong-Myung;Choi, Cha-Yong
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.11 no.3
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    • pp.258-261
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    • 2006
  • The addition of zeolite particles enhances the stability of mRNA molecules in a cell-free protein synthesis system. When $20{\mu}g/{\mu}L$ of zeolite (Y5.4) is added to a reaction mixture of cell-free protein synthesis, a substantial increase in protein synthesis is observed. The stabilizing effect of zeolite is most dearly observed in an in vitro translation reaction directed by purified mRNA, as opposed to a coupled transcription and translation reaction. Upon the addition of zeolite in the in vitro translation reaction, the life span of the mRNA molecules is substantially extended, leading to an 80% increase in protein synthesis. The effect of zeolite upon the mRNA stability appears be strongly related to the cation exchange (potassium to sodium) reaction. Our results demonstrate the possibility of modifying this biological process using heterogeneous, non-biological substances in a cell-free protein synthesis system.

Effect of Vitamin C, Silicon and Iron on Collagen Synthesis and Break-Down Enzyme Expression in the Human Dermal Fibroblast Cell (HS27) (피부 섬유아세포에서 비타민 C, Silicon, 철분 처리가 콜라겐 합성 및 분해 관련 효소의 발현에 미치는 효과 비교)

  • Kim, Jeong-Eun;Lee, Jin-Ah;Kim, Hyun-Ae;Kim, Jung-Min;Cho, Yun-Hi
    • Journal of Nutrition and Health
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    • v.42 no.6
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    • pp.505-515
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    • 2009
  • Collagen is the major matrix protein in dermis and consists of proline and lysine, which are hydroxylated by prolyl hydroxylase (PH) and lysyl hydroxylase (LH) with cofactors such as vitamin C, oxygen, iron (Fe$^{2+}$), ketoglutarate and silicon. The collagen degradation is regulated by matrix metalloproteinase-1 (MMP-1), of which is the major collagen-degrading proteinase whereas tissue inhibitors of metalloproteinase-1 (TIMP-1) bind to MMP-1 thereby inhibiting MMP-1 activity. In this study, we investigated the effects of vitamin C, silicon and iron on mRNA, protein expressions of PH, LH, MMP-1 and TIMP-1. The physiological concentrations of vitamin C (0-100 $\mu$M), silicon (0-50 $\mu$M) and iron (Fe$^{2+}$:0-50 $\mu$M) were treated to human dermal fibroblast cells (HS27 cells) for 3 or 5days. The expression level of mRNA and protein was increased in not only PH but also LH when cells were incubated with vitamin C. A similar increase in LH mRNA or protein expression occurred when cells were incubated with silicon. Our results suggest that treatment of vitamin C and silicon increased mRNA and protein expression of PH and LH in human dermal fibroblast.

Effects of Overexpression of C5 Protein on rnpB Gene Expression in Escherichia coli

  • Kim, Yool;Lee, Young-Hoon
    • Bulletin of the Korean Chemical Society
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    • v.30 no.4
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    • pp.791-793
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    • 2009
  • Escherichia coli RNase P is composed of a large RNA subunit (M1 RNA) and a small protein subunit (C5 protein). Since both subunits are assembled in a 1:1 ratio, expression of M1 RNA and C5 protein should be coordinately regulated for RNase P to be efficiently synthesized in the cell. However, it is not known yet how the coordination occurs. In this study, we investigated how overexpression of C5 protein affects expression of the rnpB gene encoding M1 RNA, using a lysogenic strain, which carries an rnpB-lacZ transcription fusion. Primer extension analysis of rnpB-lacZ fusion transcripts showed that the overexpression of C5 protein increased the amount of the fusion transcripts, suggesting that rnpB expression increases with the increase of intracellular level of C5 protein.

M Protein from Dengue virus oligomerizes to pentameric channel protein: in silico analysis study

  • Ayesha Zeba;Kanagaraj Sekar;Anjali Ganjiwale
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.41.1-41.11
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    • 2023
  • The Dengue virus M protein is a 75 amino acid polypeptide with two helical transmembranes (TM). The TM domain oligomerizes to form an ion channel, facilitating viral release from the host cells. The M protein has a critical role in the virus entry and life cycle, making it a potent drug target. The oligomerization of the monomeric protein was studied using ab initio modeling and molecular dynamics simulation in an implicit membrane environment. The representative structures obtained showed pentamer as the most stable oligomeric state, resembling an ion channel. Glutamic acid, threonine, serine, tryptophan, alanine, isoleucine form the pore-lining residues of the pentameric channel, conferring an overall negative charge to the channel with approximate length of 51.9 Å. Residue interaction analysis for M protein shows that Ala94, Leu95, Ser112, Glu124, and Phe155 are the central hub residues representing the physicochemical interactions between domains. The virtual screening with 165 different ion channel inhibitors from the ion channel library shows monovalent ion channel blockers, namely lumacaftor, glipizide, gliquidone, glisoxepide, and azelnidipine to be the inhibitors with high docking scores. Understanding the three-dimensional structure of M protein will help design therapeutics and vaccines for Dengue infection.

Action of Protein Kinase A and C Activators on Germinal Vesicle Breakdown and One-Cell Embryos in the Mouse (생쥐 GV난자와 1-세포기 배아의 핵막붕괴에 미치는 Protein Kinase A와 C의 작용)

  • 이대기;김경진;조완규
    • The Korean Journal of Zoology
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    • v.32 no.2
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    • pp.153-162
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    • 1989
  • Expedments were perfonned to examine the role of cAMP-dependent protein kinase (PK-A) and diacylglycerol-dependent protein kinase (PK-C) during the meiodc resumption and the first mitotic cell cycle of mouse embryogenesis. Mejoric GV oocytes and one-cell embryos derived from in vitro fertilization were cultured in vitro, and morphological changes in response to activators of PK-A and PK-C were examined. Treatments with a membrane-permeable cAMP analog, dbcAMP (0.1 mg/mi), phosphodiesterase inhibitor, IBMX (0.1 mM), biologically active phorbol ester, WA (10 nglmi), or a synthetic diacylglycerol, sn-diC8 inhibited resumption of melosis. Combination of PK-A and PK-C activator brought about furiher inhibition. On the contrary, dbcAMP (0.1 mg/mi), IBMX (0.2 mM), WA (10 nglml), and sn-diC8 (0.5 mM) did not inhibit pronucleus membrane breakdown (PNBD) when added S or G2 phase of cell cycle. However, activators of PK-C inhibited cleavage of one-cefl embryos. This result indicates that the action mechanism of PK-A and PK-C on dissolution of nuclear membrane in primary meiotic arrest oocytes may be different from that of mitotic one-cell embryos.

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The RecA-like protein of Schizosoccharomvces pombe: its cellular level is induced by DNA-damaging agents (DNA 상해요인에 의한 Schizosaccharomyces pombe RecA 유사 단백질의 유도생성)

  • 이정섭;박상대
    • The Korean Journal of Zoology
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    • v.37 no.2
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    • pp.232-239
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    • 1994
  • RecA protein plans a central role in homologous recombination and DNA repair in Escherichia cofi (E. colD. The function 8nd structure of this protein are universal in prokarvotes and also conserved in eukaryotes such as yeast. The RecA-like protein with 74 lInDa in size has already been identified and purified from a fission yeast Schizosaccharomyces pombe (5. pommel (Lee, 19911. From this study it was revealed that the RecA-like protein of 5. pombe was highly inducible to various DNA damaging agents and inhibitors of nucleotide pool svnthesizins enzymes. The cellular level of the 5. pombe RecA-like protein wi,u markedly increased, upto 5- to 10-fold, by treatment with various DNA-damains agents including ultraviolet (UV) light, methyl methanesulfonate WS),4-nitroquinoline-1-oxide (4-NQO), and mitomycin-C (MMC), similar to E. cofi RecA protein. Interestingly, the protein level was also increased by inhibitors of nucleotide pool forming enzlwnes such as methotrexate (MTX) and hvdroxvurea (HU). The most effective doses for the inducibility of 4-NQO, MMS, W, MMC, MTX, and HU were 0.2 Ug/ml, 30 mM, 200 J/ma, 0.4 $\mus/ml,$ 1 Ug/ml, and 100 mM, respectively. The range of effective duration time for the inducibilitv of RecA-like protein was from 270 to 450 mins. These results suggest that the 5. pombe RecA-like protein also platys an imortant role in cellular responses to DNA damage as in E. coli system.

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Influenza M1 Virus-Like Particles Consisting of Toxoplasma gondii Rhoptry Protein 4

  • Lee, Su-Hwa;Lee, Dong-Hun;Piao, Ying;Moon, Eun-Kyung;Quan, Fu-Shi
    • Parasites, Hosts and Diseases
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    • v.55 no.2
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    • pp.143-148
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    • 2017
  • Toxoplasma gondii infections occur throughout the world, and efforts are needed to develop various vaccine candidates expressing recombinant protein antigens. In this study, influenza matrix protein (M1) virus-like particles (VLPs) consisting of T. gondii rhoptry antigen 4 (ROP4 protein) were generated using baculovirus (rBV) expression system. Recombinant ROP4 protein with influenza M1 were cloned and expressed in rBV. SF9 insect cells were coinfected with recombinant rBVs expressing T. gondii ROP4 and influenza M1. As the results, influenza M1 VLPs showed spherical shapes, and T. gondii ROP4 protein exhibited as spikes on VLP surface under transmission electron microscopy (TEM). The M1 VLPs resemble virions in morphology and size. We found that M1 VLPs reacted with antibody from T. gondii-infected mice by western blot and ELISA. This study demonstrated that T. gondii ROP4 protein can be expressed on the surface of influenza M1 VLPs and the M1 VLPs containing T. gondii ROP4 reacted with T. gondii-infected sera, indicating the possibility that M1 VLPs could be used as a coating antigen for diagnostic and/or vaccine candidate against T. gondii infection.

Purification of Thiazole- and Pyrazine-inducible Microsomal Epoxide Hydrolase: Induction of Epoxide Hydrolase-related Novel 43 kDa Protein (Thiazole 또는 Pyrazine유도성 Microsomal Epoxide Hydrolase의 순수정제: Epoxide Hydrolase-관련성 43 kDa 단백질의 유도증가)

  • Kim, Sang-Geon
    • The Korean Journal of Pharmacology
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    • v.29 no.2
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    • pp.275-282
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    • 1993
  • Liver microsomal epoxide hydrolase (mEH) is active in the detoxification of epoxide-containing reactive intermediate. Previous studies in this laboratory have shown that thiazole and pyrazine are efficacious inducers of mEH in rats with large increases in mEH mRNA levels (Carcinogensis, Kim et al, 1993). mEH was purified to electrophoretic homogeneity from thiazole-induced rat hepatic microsomes using DEAE-cellulose column chromatography whereas another protein $({\sim}43\;kDa)$ was co-purified with mEH from pyrazine-induced rat hepatic micrsomes (200 mg/kg body weight/day, ip, 3d). The antibody raised from a rabbit against mEH protein purified from thiazole-induced rat hepatic microsomes appeared to specifically recognize mEH protein in rat hepatic microsomes, as assessed by immunoblotting analysis. Immunoblotting analyses revealed a 10- and 7-fold increase in mEH levels in the hepatic microsomes isolated from thiazole- and pyrazine-treated rats, respectively. Moreover, immunoblotting analysis showed cross-reactivity of the mEH antibody with a 43 kDa protein in pyrazine-induced rat hepatic microsomes and with co-purified 43 kDa protein in purified fractions. The ratio between the 43 kDa protein and mEH in pyrazine-induced rat microsomes or in purified fractions was ${\sim}1$ to 15. N-terminal amino acid sequence analysis of both purified rat mEH and 43 kDa protein revealed that 10 out of 12 amino acids in N-terminus of the 43 kDa protein were identical with the mEH sequence with two amino acid residues of the 43 kDa protein undetermined. Either thiazole or pyrazine treatment, however, failed to increase the levels of mEH protein in rabbits while pyrazine caused elevation of the 43 kDa protein in this species, as determined by irnrnunoblotting analysis. These results demonstrated that treatment of rats with either thiazole or pyrazine causes elevation in hepatic mEH expiession whereas pyrazine treatment results in induction of another mEH-related 43 kDa protein and that a distinct species difference exists between rats and rabbits in the induction of mEH by these xenobiotics.

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The Involvement of Protein Kinase C and Tyrosine Kinase in Vanadate-induced Contraction

  • Sim, Sang-Soo;Kim, Chang-Jong
    • Archives of Pharmacal Research
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    • v.21 no.3
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    • pp.315-319
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    • 1998
  • Gastric smooth muscle of cats was used to investigate the involvement of protein kinase in vanadate-induced contraction. Vanadate caused a contraction of cat gastric smooth muscle in a dose-dependent manner. Vanadate-induced contraction was totally inhibited by 2 mM EGTA and 1.5 mM $LACI_3$ and significantly inhibited by $10\mu$M verapamil and $1\mu$M nifedipine, suggesting that vanadate-induced contraction is dependent on the extracellular $Ca^{2+}$ concentration, and the influx of extracellular $Ca^{2+}$ was mediated through voltage-dependent $Ca^{2+}$ channel. Both protein kinase C inhibitor and tyrosine kinase inhibitor significantly inhibited the vanadate-induced contraction and the combined inhibitory effect of two protein kinase inhibitors was greater than that of each one. But calmodulin antagonists did not have any influence on the vanadate-induced contraction. On the other hand, both forskolin ($1\mu$M) and sodium nitroprusside ($1\mu$M) significantly inhibited vanadate-induced contraction. Therefore, these results suggest that both protein kinase C and tyrosino kinase are involved in the vanadate-induced contraction which required the influx of extracellular $Ca^{2+}$ in cat gastric smooth muscle, and that the contractile mechanism of vanadate may be different from that of agonist binding to its specific receptor.

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