• Title/Summary/Keyword: T protein

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Structural Studies on the E. coli Methionyl-tRNA Synthetase and Their Interaction with E. coli $tRNA^{fMet}$

  • Kim Ji-Hun;Ahn Hee-Chul;Park Sung-Jin;Kim Sung-Hoon;Lee Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.9 no.2
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    • pp.110-121
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    • 2005
  • E.coli methionyl tRNA synthetase consist of 676 amino acids and plays a key role in initiation of protein synthesis. The native form of this enzyme is a homodimer, but the monomeric enzyme truncated approximately C-terminal 120 amino acids retains the full enzymatic activities. X-ray crystal structure of the active monomeric enzyme shows that it has two domains. The N-terminal domain is thought to be a binding site for acceptor stem of tRNA, ATP, and methionine. The C-terminal domain is mainly a-helical and makes an interaction with the anticodon of $tRNA^{Met}$. Especially it is suggested that the region of helix-loop-helix including the tryptophan residue at the position 461 may be the essential for the interaction with anticodon of $tRNA^{Met}$. In this work the structure and function of E. coli methionyl-tRNA synthetase was studied by spectroscopic method (NMR, CD, Fluorescence). The importance of tryptophan residue at the position 461 was investigated by fluorescence spectroscopy. Tryptophan 461 is expected to be an essential site for the interaction between E. coli methionyl-tRNA synthetase and E. coli $tRNA^{Met}$. Proton and heteonuclear 2-dimensional NMR spectroscopy were also used to elucidate the protein-tRNA interaction.

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Leucaena Seeds as Protein Supplement in the Rations of Growing Sheep

  • Singh, Sultan;Kundu, S.S.;Negi, A.S.;Gupta, S.K.;Singh, N.P.;Pachouri, V.C.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.10
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    • pp.1433-1438
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    • 2002
  • The study was carried out to evaluate leucaena seeds as a protein replacement of mustard seed cake (MSC) in the concentrate mixture of growing lambs. Fifteen owing male lambs (Local${\times}$Corridale) with an average body weight of 16.3 kg were allocated into three dietary treatments (T1, T2, and T3) with five animals in each group. Animals were offered dry mixed grass, berseem hay and concentrate mixture to meet their nutrient requirements. In concentrate mixture of T1, (Control) MSC was used as protein source, while in T2 and T3 groups, 25 and 50% of MSC was replaced by leucaena leucocephala seeds. On completion of three months (90 days) of feeding, a digestion cum-metabolism trial was conducted to determine DMI, nutrient utilization, and nitrogen balance. Changes in body weight were recorded at 15 day internals and eating patterns were recorded for 3 consecutive days at the end of the feeding trial. MSC had higher CP contents than leucaena seeds (27.0%). Mimosine contents in leucaena seeds were 1.1 compared to 0.2 and 0.4% in concentrate mixture of T2 and T3 group, respectively. Dry matter intake varied non-significantly ($79.3{\pm}1.2$ to $83.4{\pm}1.3g/kg$ $w^{0.75}$) across the dietary treatments. Digestibility of DM and cell wall polysaccharides (NDF, ADF. Cellulose and hemicellulose) were comparable, however CP digestibility was relatively lower in leucaena luecocephala seeds based groups (T2 $45.5{\pm}1.7$ and T3 $46.7{\pm}3.5$) compared to MSC supplemented group (T1 $47.7{\pm}0.9%$). The growth rate of lambs was non-significantly higher in T1 ($79.2{\pm}5.4$) compared to T2 ($73.8{\pm}8.8$) and T3 ($73.9{\pm}7.0$), respectively. The animals were in positive nitrogen balance and N-balance varied from 1.8 to 2.9 g/d across treatment groups. The eating rate (% of total offered) of concentrate up-to 15 min was relatively higher in T1 (82.4) than T2 (74.2) and T3 (77.8%). However no effect of leucaena seeds was recorded on total DMI of animals. The results of the study revealed that the inclusion of up to 50% leucaena seeds, as protein source in concentrate mixture of lambs had no adverse effect on DMI, nutrient utilization, eating patterns, nitrogen balance and growth performance of lambs.

Lupeol Improves TNF-α Induced Insulin Resistance by Downregulating the Serine Phosphorylation of Insulin Receptor Substrate 1 in 3T3-L1 Adipocytes (3T3-L1 지방세포에서 루페올의 IRS-1의 인산화 조절을 통한 TNF-α 유도 인슐린 저항성 개선 효과)

  • Hyun Ah Lee;Ji Sook Han
    • Journal of Life Science
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    • v.33 no.11
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    • pp.859-867
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    • 2023
  • Lupeol is a type of pentacyclic triterpene that has been reported to have therapeutic effects for treating many diseases; however, its effect on insulin resistance is unclear clear. This study examined the inhibitory effect of lupeol on the serine phosphorylation of insulin receptor substrate-1 in insulin resistance-induced 3T3-L1 adipocytes. 3T3-L1 cells were cultured and treated with tumor necrosis factor-α (TNF-α) for 24 hours to induce insulin resistance. Cells treated with different concentrations of lupeol (15 μM or 30 μM) or 100 nM of rosiglitazone were incubated. Then, lysed cells underwent western blotting. Lupeol exhibited a positive effect on the negative regulator of insulin signaling and inflammation-activated protein kinase caused by TNF-α in adipocytes. Lupeol inhibited the activation of protein tyrosine phosphatase-1B (PTP-1B)-a negative regulator of insulin signaling-and c-Jun N-terminal kinase (JNK); it was also an inhibitor of nuclear factor kappa-B kinase (IKK) and inflammation-activated protein kinases. In addition, Lupeol downregulated serine phosphorylation and upregulated tyrosine phosphorylation in insulin receptor substrate-1. Then, the downregulated phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT) pathway was activated, the translocation of glucose transporter type 4 was stimulated to the cell membrane, and intracellular glucose uptake increased in the insulin resistance-induced 3T3-L1 adipocytes. Lupeol may improve TNF-α-induced insulin resistance by downregulating the serine phosphorylation of insulin receptor substrate 1 by inhibiting negative regulators of insulin signaling and inflammation-activated protein kinases in 3T3-L1 adipocytes.

Activities of E6 Protein of Human Papillomavirus 16 Asian Variant on miR-21 Up-regulation and Expression of Human Immune Response Genes

  • Chopjitt, Peechanika;Pientong, Chamsai;Bumrungthai, Sureewan;Kongyingyoes, Bunkerd;Ekalaksananan, Tipaya
    • Asian Pacific Journal of Cancer Prevention
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    • v.16 no.9
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    • pp.3961-3968
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    • 2015
  • Background: Variants of human papillomavirus (HPV) show more oncogenicity than do prototypes. The HPV16 Asian variant (HPV16As) plays a major role in cervical cancer of Asian populations. Some amino acid changes in the E6 protein of HPV16 variants affect E6 functions such as p53 interaction and host immune surveillance. This study aimed to investigate activities of HPV16As E6 protein on modulation of expression of miRNA-21 as well as interferon regulatory factors (IRFs) 1, 3, 7 and c-fos. Materials and Methods: Vectors expressing E6 protein of HPV16As (E6D25E) or HPV16 prototype (E6Pro) were constructed and transfected into C33A cells. HCK1T cells expressing E6D25E or E6Pro were established by transducing retrovirus-containing E6D25E or 16E6Pro. The E6AP-binding activity of E6 and proliferation of the transfected C33A cells were determined. MiR-21 and mRNA of interesting genes were detected in the transfected C33A cells and/or the HCK1T cells, with or without treatment by culture medium from HeLa cells (HeLa-CM). Results: E6D25E showed binding activity with E6AP similar to that of E6Pro. Interestingly, E6D25E showed a higher activity of miR-21 induction than did E6Pro in C33A cells expressing E6 protein. This result was similar to the HCK1T cells expressing E6 protein, with HeLa-CM treatment. The miR-21 up-regulation significantly corresponded to its target expression. Different levels of expression of IRFs were also observed in the HCK1T cells expressing E6 protein. Interestingly, when treated with HeLa-CM, IRFs 1, 3 and 7 as well as c-fos were significantly suppressed in the HCK1T cells expressing E6D25E, whereas those in the HCK1T cells expressing E6Pro were induced. A similar situation was seen for IFN-${\alpha}$ and IFN-${\beta}$. Conclusions: E6D25E of the HPV16As variant differed from the E6 prototype in its activities on epigenetic modulation and immune surveillance and this might be a key factor for the important role of this variant in cervical cancer progression.

WHOLE-BODY PROTEIN TURNOVER IN GOATS ENHANCED BY SUPPLEMENTING A DIET WITH RUMEN PROTECTED METHIONINE

  • Muramatsu, T.;Hatano, T.;Ueda, Y.;Furuse, M.;Okumura, J.
    • Asian-Australasian Journal of Animal Sciences
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    • v.7 no.2
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    • pp.279-288
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    • 1994
  • Three experiments were conducted with female Japanese Saanen goats to investigate the effects of rumen protected methionine (RPMet) on N utilization and whole-body protein turnover. Whole-body leucine flux from which whole-body protein turnover rates were derived was measured by primed- continuous infusion of L-[$^{15}N$] leucine in combination with gas chromatography-mass spectrometry. Throughout the experiments RPMet was added to a diet to supply 1.5 g DL-methionine per goat per day. Irrespective of the major N sources (i.e., protein or urea) in the diet, both N deposition and whole-body protein synthesis were increased (p<0.05), and urinary N excretion was decreased (p<0.05) by supplementing with RPMet, but not by supplementing with methionine. It was concluded, therefore, that under the present experimental conditions, the RPMet supplement was efficiently bypassed to result in enhanced body protein synthesis of the goat.

Detection of Human Anti-Trypanosoma cruzi Antibody with Recombinant Fragmented Ribosomal P Protein

  • Kim, Yeong Hoon;Yang, Zhaoshou;Lee, Jihoo;Ahn, Hye-Jin;Chong, Chom-Kyu;Maricondi, Wagner;Dias, Ronaldo F.;Nam, Ho-Woo
    • Parasites, Hosts and Diseases
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    • v.57 no.4
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    • pp.435-437
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    • 2019
  • Chagas disease is caused by the protozoan parasite Trypanosoma cruzi, and is endemic in many Latin American countries. Diagnosis is based on serologic testing and the WHO recommends two or more serological tests for confirmation. Acidic ribosomal P protein of T. cruzi showed strong reactivity against positive sera of patients, and we cloned the protein after fragmenting it to enhance its antigenicity and solubility. Twelve positive sera of Chagas disease patients were reacted with the fragmented ribosomal P protein using western blot. Detection rate and density for each fragment were determined. Fragments F1R1, F1R2, and F2R1 showed 100% rate of detection, and average density scoring of 2.00, 1.67, and 2.42 from a maximum of 3.0, respectively. Therefore, the F2R1 fragment of the ribosomal P protein of T. cruzi could be a promising antigen to use in the diagnosis of Chagas disease in endemic regions with high specificity and sensitivity.

Effect of Feeding Bypass Protein with Urea Treated Grass on the Performance of Red Kandhari Calves

  • Wankhede, S.M.;Kalbande, V.H.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.7
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    • pp.970-973
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    • 2001
  • An experiment was carried out to assess the effect of feeding concentrate mixtures varying in bypass protein levels with urea-treated or untreated grass on the performance of twelve Red Kandhari calves (14 months of age and 78.15 kg body weight) for a period of 75 days. Dry grass was treated with 4 percent urea solution and ensiled for 30 days. The CP ($N{\times}6.25$) content in urea treated grass increased from 3.96 to 8.89 percent. Two iso-caloric and iso-nitrogenous concentrate mixtures (CM-I and CM-II) varying in RDP to UDP ratio viz., 65:35 and 55.45 were prepared. The calves in control group ($T_1$) were fed concentrate mixture-I with ad libitum untreated dry grass and those in experimental group ($T_2$) were fed concentrate mixture-II with ad libitum urea treated dry grass. The dry matter consumption in group $T_2$ was significantly (p<0.01) higher as compared to group $T_1$. The total DMI in $T_1$ and $T_2$ was 146.92 and 166.95 kg respectively, whereas the DMI per day and per 100 kg body weight was 1.94 and 2.22 and 1.90 and 2.35 kg, respectively. The average total gain in body weight (kg) and average daily gain (g) of calves in $T_2$ was significantly (p<0.01) higher as compared to those in $T_1$ the values being 28.66, 18.33 and 382.16, 244.44, respectively. Feed efficiency in terms of kg DM per kg gain in body weight was significantly (p<0.01) lower in group $T_1$ than in $T_2$. The cost of feed per kg gain in body weight for $T_2$ and $T_1$ group was Rs. 21.14, 28.22, respectively. The digestibility coefficients of DM, CP, EE, CF, NFE, NDF and ADF were 59.60, 57.50, 53.00, 65.04, 45.82, 48.48, 52.48 and 55.73 for $T_1$ group. The coressponding values were 68.78, 67.80, 59.83, 71.41, 49.93, 53.37 and 57.81, respectively for $T_2$ group. The digestibility coefficients for all the proximate principles in $T_2$ were significantly (p<0.01) higher as compared to $T_1$. However, NDF and ADF digestibilities were not significantly different. Nutritive value determined in terms of DCP and TDN for The experimental ration was significantly (p<0.01) higher than control ration, the values being 7.32 and 47.34 and 9.39 and 52.40% respectively. The blood urea nitrogen levels at 0, 3 and 6 h interval after feeding were significantly (p<0.01) lower in calves fed experiment ration as compared to control. The overall results indicated that in Red Kandhari calves an optimum growth can be economically achieved by feeding 4 percent urea treated dry and mature grass as basal roughage supplemented with a concentrate mixture containing 20 percent CP, 70% TDN and 45% UDP/bypass protein.

Nutrient intake, digestibility and performance of Gaddi kids supplemented with tea seed or tea seed saponin extract

  • Kumar, M.;Kannan, A.;Bhar, R.;Gulati, A.;Gaurav, A.;Sharma, V.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.4
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    • pp.486-494
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    • 2017
  • Objective: An experiment was conducted to determine the nutrient intake, digestibility, microbial protein synthesis, haemato-biochemical attributes, immune response and growth performance of Gaddi kids fed with oat fodder based basal diet supplemented with either tea seed or tea seed saponin (TSS) extract. Methods: Eighteen male kids, $7.03{\pm}0.16$ months of age and $19.72{\pm}0.64kg$ body weight, were distributed into three groups, $T_0$ (control), $T_1$, and $T_2$, consisting of 6 animals each in a completely randomized design. The kids were fed a basal diet consisting of concentrate mixture and oat fodder (50:50). Animals in group III ($T_2$) were supplemented with TSS at 0.4% of dry matter intake (DMI), and group II ($T_1$) were supplemented with tea seed at 2.6% of DMI to provide equivalent dose of TSS as in $T_2$. Two metabolism trials were conducted, 1st after 21 days and 2nd after 90 days of feeding to evaluate the short term and long term effects of supplementation. Results: The tea seed ($T_1$) or TSS ($T_2$) supplementation did not affect DMI as well as the digestibility of dry matter, organic matter, crude protein, neutral detergent fibre, and acid detergent fibre. Nutritive value of diet and plane of nutrition were also comparable for both the periods. However, the average daily gain and feed conversion ratio (FCR) were improved (p<0.05) for $T_1$ and $T_2$ as compared to $T_0$. The microbial protein supply was also higher (p<0.05) for $T_1$ and $T_2$ for both the periods. There was no effect of supplementation on most blood parameters. However, the triglyceride and low density lipoprotein cholesterol levels decreased (p<0.05) and high density lipoprotein-cholesterol level increased (p<0.05) in $T_2$ as compared with $T_0$ and $T_1$. Supplementation also did not affect the cell mediated and humoral immune response in goats. Conclusion: Tea seed at 2.6% of DMI and TSS at 0.4% DMI can be fed to Gaddi goats to improve growth rate, FCR and microbial protein synthesis.

Effect of Temperature on Mitotic Cycle of Rice Root Meristem Cells (벼 뿌리세포의 유사분열주기에 대한 배양온도의 영향)

  • 김재철;이승준;권성환;곽성희
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.35 no.1
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    • pp.65-72
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    • 1990
  • The mitotic cycle duration and component plase periods of rice (Oryza sativa L. 'Sumjinbyeo and Seokwangbyeo') root merisem cells at 15, 20, and 30$^{\circ}C$ were determined the use of tritiated thymidine, In this work, the time interval between the maxima of sequential mitotic appearances of marked cells was used to estimate the mitotic cycle duration (MCD) of rice, The MCD of rice of the cultivar 'Sumjinbyeo' and 'Seokwangbyeo' at 20. and 30$^{\circ}C$ was 12. and 20hr, respectively. But the MCD of 'Sumjinbyeo' and 'Seokwangbyeo' was 18 and 20hr, at 15$^{\circ}C$. respectively. The MCD decreased with increasing temperature, The duration of component phase of rice cultivar 'Seokwangbyeo' were essentially the same ratio at 20$^{\circ}C$ and 30$^{\circ}C$. but in 'Sumjinbyeo' cultivar the ratio of $G_1$ period was almost doubled while those of $G_2$ and M were decreased by almost two times at 20$^{\circ}C$ and 30$^{\circ}C$. Deoxyribonucleic acid (DNA). Ribonucleic acid (RNA), and protein synthesis were reduced with increasing temperature from 15$^{\circ}C$ to 30$^{\circ}C$ while the MCD was decreased, This result suggest that DNA, RNA and protein synthesis may not affect the MCD from 15$^{\circ}C$ to 30$^{\circ}C$ in rice.

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EFFECTS OF MACROPHAGE INFLAMMATORY $PROTEIN-1{\alpha}$ON THE T CELL PROLIFERATION AND THE EXPRESSION OF CD4 AND CD8 (Macrophage Inflammatory Protein $1{\alpha}$가 T세포성장 및 CD4, CD8 발현에 미치는 영향)

  • Choi, Jong-Sun;Kim, Oh-Whan
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.18 no.1
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    • pp.153-163
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    • 1996
  • Macrophage inflammatory protein $(MIP)-1{\alpha}$ is a cytokine which produces wide range of bioactivities such as proinflammatory, immunomodulatory, and hematopoietic modulatory actions. To determine whether $MIP-1{\alpha}$ acts as a negative regulator on the functions of lymphocyte, $[^3H]$-thymidine incorporation test and flow cytometric analysis were performed by using human tonsil T cell, human peripheral blood T cell, and murine cytolytic T lymphocyte (CTL) line CTLL-2, The results were as follow. 1. When human tonsil T lymphocytes were stimulated with anti-CD3 monoclonal antibody (mAb), rate of T cell proliferation was about four times increased. 200ng/ml of $MIP-1{\alpha}$ inhibited anti-CD3 mAb-mediated T cell growth as much as 60% (P<0.05). 2. The suppression of human peripheral T cell proliferation produced by $MIP-1{\alpha}$ was dramatic, but variable among T cells derived from different individuals $(40%{\sim}90%)$. 3. $MIP-1{\alpha}$inhibited the proliferation of murine CTL line CTLL-2 as much as 75%(P<0.001). 4. When the $MIP-1{\alpha}$ was added to human peripheral T cell, cell proporation of $CD4^+$ helper T cell and $CD8^+$ CTL were not noticeably affected. The expression level of CD4, not of Cd8, however, was down regulated by $MIP-1{\alpha}$ treatment $(27%{\sim}82%)$.

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