• Title/Summary/Keyword: Protein X

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Characterization and Partial Purification of Storage Protein-i Receptor in the Fat Body of Hyphantria cunea (미국흰불나방 지방체에서 저장단백질-1의 수용체의 특성과 부분정제)

  • 채권석;여성문;김학렬
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.490-497
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    • 1995
  • In vitro tissue culture of fat body of Hyphantria cunea in the medium containing [35S]-methionine reveaied that storage protein-i (SP-1) is taken up into fat body of prepupae and 1-day-old pupae. Using Western blotting and ligand binding method, we were able to identify the protein band of the SP-1 receptor protein. For the partial purification, the membrane proteins of fat body cells were solubilixed with 1% Triton X-1OO and applied to anion exchange chromatography. The results revealed the molecular weight of the receptor protein to be about 80 kl)a in SDSPAGE, and the P1 was estimated to be about 6.1. The mobility of the receptor protein in 8D8-PAGE was highly dependent on both temperature during electrophoresls and the condition of samples whether they were in reducing or nonreducing.

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Characterization of physiochemical and nutrient profiles in canola feedstocks and co-products from bio-oil processing: impacted by source origin

  • Alessandra M. R. C. B. de Oliveira;Peiqiang Yu
    • Animal Bioscience
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    • v.36 no.7
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    • pp.1044-1058
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    • 2023
  • Objective: The objective of this study was to characterize physiochemical and nutrient profiles of feedstock and co-products from canola bio-oil processing that were impacted by source origin. The feedstocks and co-products (mash, pellet) were randomly collected from five different bio-oil processing plants with five different batches of samples in each bio-processing plant in Canada (CA) and China (CH). Methods: The detailed chemical composition, energy profile, total digestible nutrient (TDN), protein and carbohydrate subfractions, and their degradation and digestion (CNCPS6.5) were determined. Results: The results showed that TDN1x was similar in meals between CA and CH. CH meals and feedstock had higher, truly digestible crude protein (tdCP) and neutral detergent fiber (tdNDF) than CA while CA had higher truly digestible non-fiber carbohydrate (tdNFC). The metabolizable energy (ME3x), net energy (NELp3x, NEm3x, and NEg3x) were similar in meals between CA and CH. No differences were observed in energy profile of seeds between CA and CH. The protein and carbohydrate subfractions of seeds within CH were similar. The results also showed that pelleting of meals affected protein sub-fractionation of CA meals, except rapidly degradable fractions (PB1), rumen degradable (RDPB1) and undegrdable PB1 (RUPB1), and intestinal digestible PB1 (DIGPB1). Canola meals were different in the soluble (PA2) and slowly degradable fractions (PB2) between CA and CH. The carbohydrate fractions of intermediately degradable fraction (CB2), slowly degradable fraction (CB3), and undegradable fraction (CC) were different among CH meals. CH presented higher soluble carbohydrate (CA4) and lower CB2, and CC than CA meals. Conclusion: The results indicated that although the seeds were similar within and between CA and CH, either oil-extraction process or meal pelleting seemed to have generated significantly different aspects in physiochemical and nutrient profiles in the meals. Nutritionists and producers need to regularly check nutritional value of meal mash and pellets for precision feeding.

In vitro Folding of Recombinant Hepatitis B Virus X-Protein Produced in Escherichia coli: Formation of Folding Intermediates

  • Kim, Sun-Ok;Sohn, Mi-Jin;Jeong, Soon-Seog;Shin, Jeh-Hoon;Lee, Young-Ik
    • BMB Reports
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    • v.32 no.6
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    • pp.521-528
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    • 1999
  • The folding of recombinant hepatitis B virus X-protein (rHBx) solubilized from Escherichia coli inclusion bodies was investigated. By sequential dialysis of urea, rHBx was folded into its native structure, which was demonstrated by the efficacy of its transcriptional activation of the adenovirus major late promoter (MLP), fluorescence spectroscopy, and circular dichroism (CD) analysis. The decrease in CD values at 220 nm and a corresponding blue shift of the intrinsic fluorescence emission confirmed the ability of rHBx to refold in lower concentrations of urea, yielding the active protein. Equilibrium and kinetic studies of the refolding of rHBx were carried out by tryptophan fluorescence measurements. From the biphasic nature of the fluorescence curves, the existence of stable intermediate states in the renaturation process was inferred. Reverse phase-high performance liquid chromatography (RP-HPLC) analysis further demonstrated the existence of these intermediates and their apparent compactness.

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Hypothermia Regulates Endoplasmic Reticulum (ER) Stress through the X-box Binding Protein-1 (XBP1) Gene Expression in PC12 Cells

  • Yoo, Bo-Kyung;Kwon, Kisang;Lee, Eun Ryeong;Kwon, O-Yu
    • Biomedical Science Letters
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    • v.23 no.4
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    • pp.416-420
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    • 2017
  • Endoplasmic reticulum (ER) stress induces unfolded protein response (UPR) via inositol-requiring enzyme 1 (IRE1) activation, which sends a molecular signal for X box-binding protein 1 (XBP1) mRNA splicing in the cytosol. IRE1 endoribonuclease activity induces cleavage of XBP1 mRNA. The XBP1 mRNA is then ligated by an uncharacterized RNA ligase and translated to produce spliced XBP1 by 23 nt removed in which contains the PstI restriction enzyme site. The splicing of XBP1 mRNA can be detected by semiquantitative RT-PCR, and then splicing of XBP1 is a useful tool to measure the genetic variability in ER stress. In this study, we have estimated IRE1-dependent splicing of XBP1 mRNA under conditions of various hypothermia. The results indicated that hypothermia regulated ER stress. This study demonstrated that hypothermia is closely related to ER stress and may be useful for early diagnosis of ER-associated disease.

Effect of Ginseng Saponins on $K^+-Dependent$ Phosphatase Activity of Dog Cardiac Sarcolemma (인삼 사포닌이 개 심실 형질막의 $K^+$-의존성 포스파타제 활성에 미치는 영향)

  • Lee, Shin-Woong;Lee, Jeung-Soo
    • YAKHAK HOEJI
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    • v.36 no.2
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    • pp.129-136
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    • 1992
  • The effects of ginseng saponins, gypsophila saponin, sodium dodecyl sulfate(SDS), and Triton X-100 on membrane $K^+-dependent$ phosphatase activity which is lipid dependent and represents dephosphorylation step of the complete Na+, $K^+-ATPase$ reaction were investigated in this study to elucidate whether the effects of ginseng saponins are due to the detergent action, using sarcolemma enriched preparation isolated from dog ventricle. $Na^+$, $K^+-ATPase$ and $K^+-dependent$ phosphatase activities of cardiac sarcolemma were about $143\;{\mu}mol$ Pi/mg protein/hr and $34\;{\mu}mol$ p-nitrophenol/mg protein/hr, respectively. While ginseng saponins (triol>total>diol) inhibited $K^+-dependent$ phosphatase activity, gypsophila saponin, and low dose of SDS($0.4\;{\mu}g/{\mu}g$ protein), and Triton X-100 ($0.6\;{\mu}g/{\mu}g$ protein) increased the enzyme activity, indicating disruptive effect of detergents on membrane barriers. The activating effect of low doses of Triton X-100 on membrane $K^+-dependent$ phosphatase appeared at concentration decreasing light scattering. However, the inhibitory effect of ginseng saponin appeared before a decrease in light scattering. These results suggest that low concentrations of ginseng saponins inhibit the membrane $K^+-dependent$ phosphatase by interacting directly with enzyme before membrane disruption.

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Effect of Water Temperature on the Expression of Stress Related Genes in Atlantic Salmon (Salmo salar) Fry (수온이 대서양 연어(Salmo salar) 치어의 체내 스트레스 관련 유전자 발현에 미치는 영향)

  • Kang, Hee Woong;Kim, Kwang Il;Lim, Hyun Jeong;Kang, Han Seung
    • Korean Journal of Environmental Biology
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    • v.36 no.2
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    • pp.131-139
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    • 2018
  • The warming of water as a result of climate change affects fish habitat. Variations in water temperature affect fish physiology almost totally. The rise in water temperature due to climate change leads to hypoxia following decreased oxygen solubility and decreased binding capacity of oxygen-carrying hemoglobin. This study was conducted to evaluate the health status of Atlantic salmon (Salmo salar) fry at elevated water temperatures($20^{\circ}C$) compared with optimum water temperature ($15^{\circ}C$). The method facilitated the detection of biomarker genes using NGS RNAseq analysis and evaluation of their expression pattern using RT-qPCR analysis. The biomarker genes included interferon alpha-inducible protein 27-like protein 2A transcript variant X3, protein L-Myc-1b-like, placenta growth factor-like transcript variant X1, fibroblast growth factor receptor-like 1 transcript variant X1, transferrin, intelectin, thioredoxin-like, c-type lectin lectoxin-Thr1-like, ladderlectin-like and calponin-1. The selected biomarker genes were sensitive to changes in water temperature based on NGS RNAseq analysis. The expression patterns of these genes based on RT-qPCR were similar to those of NGS RNAseq analysis.

Effects of Reduced Glutathione and Ginseng Extract on Non-Protein Sulfhydryl, and Non-Protein Disulfide of Mouse Liver and Blood Following Whole Body X-Irradiation (Reduced glutathione 및 인삼추출액(人蔘抽出液)이 X-선전신조사(線全身照射)를 입은 마우스 간조직(肝組織) 및 혈중(血中) NP-SH 및 NP-SS에 미치는 영향(影響))

  • Oh, Jang-Suk
    • The Korean Journal of Physiology
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    • v.6 no.2
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    • pp.57-63
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    • 1972
  • In an attempt to better understand the radioprotective effect of reduced glutathione(GSH), and to observe a possible radioprotective effect of Ginseng extract, whole body X-irradiation of 1,200 r was administered to the mouse either independently or immediately following the injection of GSH or Ginseng extract to the mouse intraperitoneally. The non-protein sulfhydryl (NP-SH) and non-protein disulfide (NP-SS) levels of the liver, and NP-SH level of NP-SH of the blood of the mouse were measured at 30, 60 and 120 minutes, and results were compared with the normal. The results thus obtained are summarized as follows; 1) The normal values of NP-SH and NP-SS of the mouse liver were $5.90{\pm}0.46\;{\mu}\;mol/gm\;wet\;wt.,\;and\;3.02{\pm}0.42\;{\mu}\;mol/ml$ wet wt., respectively, and the normal value of NP-SH of NP-SH of the mouse blood was $3.98{\pm}1.29\;{\mu}\;mol/ml$ 2) The injection of both GSH and Ginseng extract produced the highest values of NP-SH in the liver at 30 minutes, but a gradual decrease to the normal was observed thereafter. When X-irradiation alone was applied, the liver NP-SH value was lower than the normal at 60 minutes post-irradiation and thereafter. When Ginseng extract was injected immediately prior to X-irradiation, the liver NP-SH was lower than the normal throughout the experiment with the lowest value at 60 minutes. However, the combination of GSH and X-irradiation produced higher than the normal values throughout the entire experiment. 3) The liver NP-SS value was most significantly elevated at 30 minutes after the injection of GSH, hut the recovery to the normal was observed thereafter. The injection of Ginseng extract produced slightly higher liver NP-SS values at 30 and 60 minutes, but the value at 120 minutes was similar to the normal. The single application of X-irradiation resulted in the lower then normal liver NP-SS values throughout the entire experiment. When GSH was injected price to X-irradiation, the liver NP-SS values were higher than the normal at 30 and 60 minutes followed b the recovery to the normal at 120 minutes. The combination of Ginseng extract and X-irradiation showed generally lower liver NP-SS values throughout the experiment. 4) The blood NP-SH showed the higher than the normal values in all the experimental groups except when GSH was injected prior to X-irradiation alone produced e significantly elevated blood NP-SS value at 30 minutes post-irradiation.

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Construction of Recombinant Xanthomonas campestris Strain Producing Insecticidal Protein of Bacillus thuringiensis

  • Shin, Byung-Sik;Koo, Bon-Tag;Choi, Soo-Keun;Park, Seung-Hwan
    • Journal of Microbiology and Biotechnology
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    • v.4 no.4
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    • pp.285-289
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    • 1994
  • An insecticidal crystal protein gene, cryIA(c), from Bacillus thuringiensis HD-73 was integrated into the chromosome of a xanthan-producing bacterium, Xanthomonas campestris XP92. The cryIA(c) gene expression cassette was constructed that placed the gene between the trc promoter and rrnB transcriptional terminator. The $lacl^q$ gene was also included to prevent the expression of cryIA(c) gene in X campestris cells. Southem blot analysis confirmed the integration of the cryIA(c) gene expression cassette in chromosome of X campestris XP92 transconjugant. Expression of the insecticidal crystal protein was confirmed by Western blot analysis and bioassay against the larvae of Hyphantria cunea (Lepidoptera: Arctiidae) and Plutella xylostella (Lepidoptera:Plutellidae).

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Metabolizable Energy Values of Some Poultry Feeds Determined by Various Methods and Their Estimation Using Metabolizability of the Dry Matter

  • Han, I. K.;Hochstetler, H.W.;Scott, M.L.
    • Korean Journal of Poultry Science
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    • v.3 no.1
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    • pp.11-18
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    • 1976
  • Metabolizable energy (M.E.) values of 12 U.S. feedstuffs and 10 Korean feed ingredients for poultry were determined both by the total collection method and by the chromic oxide indicator method. It was found that M.E. values of most poultry feedstuffs can be measured accurately by either method. Limitation of feed intake to almost maintenance level(approximately 60% of ad libitum) did not increase or decrease the M.E. value of the feeds. An attempt was made to establish a prediction equation to estimate the M.E. values based on the apparent metabolizability of dry matter (D.M.) in the feedstuffs. The results indicated that linear relationships do exist between D. M. metabolizability and M.E. values of carbohydrate-rich feedstuffs (grains and their by-products) or protein-rich feed ingredients (oil seed meals and animal protein feeds) or lipid-rich feeds (fats and oils) as follows: The prediction equation for carbohydrate-rich feedstuffs was Y = 0.0947x - 3.498 ($r^2\;=\;0.99$, Sy.x = 0.015); for protein-rich feed ingredients. it was Y = 0.1234x - 4.898 ($r^2\;=\;0.99$, Sy.x = 0.027); and for lipid-rich feedstuffs it was Y = 0.0844x + 0.774 ($r^2\;=\;0.99$, Sy.x = 0.032). where x = metabolizability of dry matter of feeds in %, and Y=metabolizable energy values in kcal./g. The errors attached to these estimations were relatively small. Thus these prediction equations may be very useful for estimation of the M.E. values from D.M. apparent metaboiizability of feeds, especially in areas of the world where calorimetry is not possible.

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Increased Stability of Nucleolar PinX1 in the Presence of TERT

  • Keo, Ponnarath;Choi, Joong Sub;Bae, Jaeman;Shim, Yhong-Hee;Oh, Bong-Kyeong
    • Molecules and Cells
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    • v.38 no.9
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    • pp.814-820
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    • 2015
  • PinX1, a nucleolar protein of 328 amino acids, inhibits telomerase activity, which leads to the shortening of telomeres. The C-terminal region of PinX1 is responsible for its nucleolar localization and binding with TERT, a catalytic component of telomerase. A fraction of TERT localizes to the nucleolus, but the role of TERT in the nucleolus is largely unknown. Here, we report a functional connection between PinX1 and TERT regarding PinX1 stability. The C-terminal of $PinX1^{205-328}$, a nucleolar fragment, was much more stable than the N-terminal of $PinX1^{1-204}$, a nuclear fragment. Interestingly, PinX1 was less stable in TERT-depleted cells and more stable in TERT-myc expressing cells. Stability assays for PinX1 truncation forms showed that both $PinX1^{1-328}$ and $PinX1^{205-328}$, nucleolar forms, were more rapidly degraded in TERT-depleted cells, while they were more stably maintained in TERT-overexpressing cells, compared to each of the controls. However, $PinX1^{1-204}$ was degraded regardless of the TERT status. These results reveal that the stability of PinX1 is maintained in nucleolus in the presence of TERT and suggest a role of TERT in the regulation of PinX1 steady-state levels.