• Title/Summary/Keyword: recovered protein

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A Thermodynamic Study on the Binding of Cobalt Ion with Myelin Basic Protein

  • Behbehani, G. Rezaei;Saboury, A.A.;Baghery, A. Fallah
    • Bulletin of the Korean Chemical Society
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    • v.29 no.4
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    • pp.736-740
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    • 2008
  • The interaction of myelin basic protein (MBP) from bovine central nervous system with divalent calcium ion was studied by isothermal titration calorimetry at 27 ${^{\circ}C}$ in aqueous solution. The extended solvation model was used to reproduce the enthalpies of $Co^{2+}$-MBP interaction over the whole $Co^{2+}$ concentrations. The solvation parameters recovered from the solvation model were attributed to the structural change of MBP due to the metal ion interaction. It was found that there is a set of three identical and noninteracting binding sites for $Co^{2+}$ ions. The association equilibrium constant is 0.015 ${\mu}M^{-1}$. The molar enthalpy of binding is $\Delta$H = −14.60 kJ $mol^{-1}$.

Cellular Distribution and Metabolism of Ginsenosides in Rat Liver (쥐 간에서의 Ginsenoside의 세포내 분포와 대사)

  • 윤수희;이희봉
    • Journal of Ginseng Research
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    • v.17 no.2
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    • pp.114-122
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    • 1993
  • 0.5 mg of natural ginsenoside mixture and 0.8 $\mu$Ci of synthesized 14C-ginsenosides were administered orally to a rat and killed at one hour after the ginsenoside administration and the liver was fractionated into nuclear fraction, mitrochondria microsomes and cytosol fraction. Radioactivity distribu lion in subcellular fractions of the liver showed that 32o1c of total radioactivity absorbed in the liver was in cytosol fraction but a significant portion of the radioactivity was also found in mitochondria (26.6%) and microsomal fraction (18.l%). 5.8% of the total radioactivity was recovered from the nuclear fraction as well. This suggested that ginsenosides might be distributed into all subcellular fractions. Activities of mitochondrial aldehyde dehydrogenase, lactate dehydrogenase and malate dehydrogenase of the liver of rat at two hours after the ginsenoside administraion were found appreciably stimulated, suggesting that the ginsenoside concentration in the liver might be around 10-5%, since optimum concentrations for most enzyme catalyzed reactions in vitro were known to be 10-6% 10-4%. A significant portion of the radioactivity recovered from subcellular fractions of the liver was found in protein fractions, suggesting that proteins might interact with ginsenosides. Examination of protein-ginsenoside interation by gel filtration, equilibrium dialysis and amonium sulfate precipitation technique suggesting that proteins and ginsenosides do not bound covalently but weakl\ulcorner combined. When purified ginsenoside Rbl and Rgl were incubated with rat liver cytosolic enzymes for 20 min, the above ginsenosides were hydrolyzed quickly, suggesting that ginsenosides might be rapidly hydrolyzed and metabolized in the liver. It was also observed in vitro that the ginsenosides such as Rbl and Rgl were easily hydrolyzed by rat liver cytosol preparation suggesting that absorbed ginsenosides might be quickly hydrolyzed and metabolized in the liver.

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Recovery of Soy Oligosaccharides using Calcium Oxide (산화칼슘을 이용한 대두 올리고당의 회수)

  • Choi, Yeon-Bae;Kim, Kang-Sung;Sohn, Heon-Soo
    • Korean Journal of Food Science and Technology
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    • v.27 no.2
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    • pp.225-229
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    • 1995
  • Soy oligosaccharide, a low calorie sugar, which is known to improve the intestinal microbial flora, was recovered from the waste of soymilk process by Steffen process. To remove protein contaminants, prior to the Steffen process, pH of the sample was adjusted to $3.5{\sim}4.0$ or calcium chloride was added 8%(w/w) per sugar. Both pretreatment processes were found to remove about $25{\sim}30%$ of the protein initially present in the sample. Using the Steffen process, as much as 85% of soy oligosaccharide could be recovered as a saccharate form. The amounts of calcium chloride and lime used were 20%(w/w) and $100{\sim}120%$(w/w) per total sugar, respectively. After the sugar was desorbed by $CO_{2}$, the final yield of oligosaccharide was 80% while 80% of protein were removed from the original solution. The composition of sugar was similar to that of soybean cooking water.

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Immunopotentiating Effect of Protein Extract from Dioscorea quinqueloba in Stressed Mice (단풍마 단백질 추출물의 스트레스로 인한 면역력 저하 개선 효과)

  • Kim, Ju Hwan;Lee, Sun-Mee;Lee, Dong-Cheol
    • The Korean Journal of Food And Nutrition
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    • v.31 no.2
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    • pp.252-257
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    • 2018
  • It is noted that Dioscorea quinqueloba is a medicinal herb that is widely used to treat cardiovascular disease and is assessed as useful to treat other various medical conditions. The immunopotentiating effects of the protein extract (DQP-1) from Dioscorea quinqueloba were thus formally investigated in vivo under incident of cold stress. In this case study, the spleen and thymus weight in mice was shown to have decreased after a measured exposure to cold stress, while the adrenal gland weight in the mice was shown to have increased. The systematic oral administration of DQP-1 significantly recovered the weight loss of the spleen and suppressed the adrenal gland hypertrophy during the association with cold stress. Additionally, the DQP-1 also restored the ascorbic acid level in the adrenal gland reduced after cold stress. The cold stress exposure lowered the percentage of $CD4^+$ and $CD8^+$ cells in the mouse thymus as determined by the flow cytometric analysis, as well as the levels of some serum immunological cytokines(interleukin-2, interleukin-12, and interferon-${\gamma}$) in the studied mice. The resulting identified weakened immunity caused by cold stress was also recovered by a treatment with DQP-1. The DQP-1 significantly suppressed the formation of serum enzymes of alanine aminotransferase, aspartate aminotransferase, and lactate dehydrogenase, which were systematically elevated during the cold stress episode. These results indicate that DQP-1 can improve immunity in mice that are characteristically weakened under stress.

Recovery and Utilization of Proteins and Lipids from the Washing Wastewater in Marine Manufacture by Isoelectric Point Shifting Precipition Method -2. Utilization of the Recovered Proteins as the Material of a Processed Food- (수산가공공장폐액의 등전점이동 응집처리에 의한 유용성분재회수이용 -2. 회수단백질의 가공식품소재로서의 이용-)

  • SUH Jae-Soo;CHO Soon-Yeong;SON Kwang-Tae;KIM Jin-Soo;LEE Eung-HO
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.5
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    • pp.495-500
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    • 1994
  • Mackerel water-soluble protein solution Mackerel meat washing water were concentrated by isoelectric point shifting precipitation process, and the concentrates were utilized as a material for processing of an elastic gel food such as kamaboko. The water-soluble proteins were partly polymerized during the isoelectric point shifting precipitation process. Then, the water soluble protein concentrates were partly substituted for frozen minced Alaska pollack meat in processing of a good quality kamaboko. The maximum substitution percentage for good quality kamaboko manufacturing was concluded to be below $30\%$, according to the criteria of color difference, jelly strength and folding tests using the substituted recovered protein concentrates.

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Introduction and Expression of a Thaumatin-like Protein from Rice in American Ginseng Following Agrobacterium-mediated Transformation

  • Chen, W.P.;Punja, Z.K.
    • Journal of Ginseng Research
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    • v.27 no.1
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    • pp.17-23
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    • 2003
  • Agrobacterium-mediated transformation of American ginseng (Panax quinquefolius L.) with strain LBA 4404 containing a rice thaumatin-like protein gene is described. The selectable markers used were phosphinothricin acetyltransferase and hygromycin phosphotransferase genes. Epicotyl explants from seedlings were precultured for 5-7 days on Murashige and Skoog medium with ${\alpha}$-naphthaleneacetic acid and 2,4 dichlorophenoxyacetic acid at 10 ${\mu}$M and 9 ${\mu}$M, respectively (ND medium), prior to Agrobacterium infection. The explants were immersed in a bacterial suspension for 20 min. A post-infection co-culture period of 3-4 days was provided on ND medium. Selection for transformed calli was conducted on ND medium with 20 mg/L phosphinothricin followed by 100 mg/L hygromycin over an 8-month period. it transformation frequency of 24.8% was achieved at the callusing phase. The presence of the transgenes in calli was confirmed by Southern hybridization and polymerase chain reaction analysis. The expression of the thaumatin-like protein gene in ginseng calli was demonstrated by Western blot analysis. Somatic embryos were produced from both transgenic calli and suspension cultures, and plantlets were recovered that expressed the transgenic thaumatin-like protein gene.

Recombinant human BMP-2/-7 heterodimer protein expression for bone tissue engineering using recombinant baculovirus expression system

  • Park, Seung-Won;Goo, Tae-Won;Kim, Seong Ryul;Choi, Kwang-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.32 no.2
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    • pp.49-53
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    • 2016
  • Bone morphogenetic proteins (BMPs) are essential growth factors for bone formation, skeletal development and bone regeneration. The BMP-2/7 heterodimer is known to have remarkable effects on osteogenic induction that are even stronger than the BMP-2 or BMP-7 homodimers. We designed a recombinant human BMP-2/7 (rhBMP-2/7) heterodimer protein with four glycine residues between BMP-2 and BMP-7 protein to facilitate free bond rotation of domains. The Baculovirus Expression Vector System (BEVS) is routinely used to produce recombinant proteins in the milligram scale. In this study, the BEVS was used to express the rhBMP-2/7 protein whrer the recombinant baculovirus was recovered in the host Sf9 cells. To confirm the biological activity of rhBMP-2/7 protein secreted from the BEVS as an osteogenic differentiation and induction factor, we measured the BMP-induced ALP activity. rhBMP-2/7 could be used as an alternative to BMPs to overcome limitations like short half-life and requirement for high concentrations. Furthermore, rhBMP-2/7 may be an efficient tool for various application studies such as bone regeneration and skeletal development.

A protein interactions map of multiple organ systems associated with COVID-19 disease

  • Bharne, Dhammapal
    • Genomics & Informatics
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    • v.19 no.2
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    • pp.14.1-14.6
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    • 2021
  • Coronavirus disease 2019 (COVID-19) is an on-going pandemic disease infecting millions of people across the globe. Recent reports of reduction in antibody levels and the re-emergence of the disease in recovered patients necessitated the understanding of the pandemic at the core level. The cases of multiple organ failures emphasized the consideration of different organ systems while managing the disease. The present study employed RNA sequencing data to determine the disease associated differentially regulated genes and their related protein interactions in several organ systems. It signified the importance of early diagnosis and treatment of the disease. A map of protein interactions of multiple organ systems was built and uncovered CAV1 and CTNNB1 as the top degree nodes. A core interactions sub-network was analyzed to identify different modules of functional significance. AR, CTNNB1, CAV1, and PIK3R1 proteins were unfolded as bridging nodes interconnecting different modules for the information flow across several pathways. The present study also highlighted some of the druggable targets to analyze in drug re-purposing strategies against the COVID-19 pandemic. Therefore, the protein interactions map and the modular interactions of the differentially regulated genes in the multiple organ systems would incline the scientists and researchers to investigate in novel therapeutics for the COVID-19 pandemic expeditiously.

USP44 Promotes the Tumorigenesis of Prostate Cancer Cells through EZH2 Protein Stabilization

  • Park, Jae Min;Lee, Jae Eun;Park, Chan Mi;Kim, Jung Hwa
    • Molecules and Cells
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    • v.42 no.1
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    • pp.17-27
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    • 2019
  • Ubiquitin-specific protease 44 (USP44) has been implicated in tumor progression and metastasis across various tumors. However, the function of USP44 in prostate cancers and regulatory mechanism of histone-modifying enzymes by USP44 in tumors is not well-understood. Here, we found that enhancer of zeste homolog 2 (EZH2), a histone H3 lysine 27 methyltransferase, is regulated by USP44. We showed that EZH2 is a novel target of USP44 and that the protein stability of EZH2 is upregulated by USP44-mediated deubiquitination. In USP44 knockdown prostate cancer cells, the EZH2 protein level and its gene silencing activity were decreased. Furthermore, USP44 knockdown inhibited the tumorigenic characteristics and cancer stem cell-like behaviors of prostate cancer cells. Inhibition of tumorigenesis caused by USP44 knockdown was recovered by ectopic introduction of EZH2. Additionally, USP44 regulates the protein stability of oncogenic EZH2 mutants. Taken together, our results suggest that USP44 promotes the tumorigenesis of prostate cancer cells partly by stabilizing EZH2 and that USP44 is a viable therapeutic target for treating EZH2-dependent cancers.

Change of Antioxidant Enzymes Activities in Leaves of Soybean(Glycine max) during Water Stresses and Following Recovery (대두에서 수분장해 및 회복시 엽중 항산화효소의 활성 변화)

  • Kang, Sang-Jae;Kim, Tae-Sung;Park, Woo-Churl
    • Korean Journal of Soil Science and Fertilizer
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    • v.32 no.2
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    • pp.164-170
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    • 1999
  • This experiment was carried out to elucidate change of antioxidant enzymes activities subjected to water stresses in soybean plant. In this study, we measured the activities of ascorbate peroxidase(APDX), monodehydroascorbate reductase(MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase(GR) subjected to drought or flooding stresses for 4days and following recovery for 3days. Leaves of two soybean lines subjected to drought or flooding showed premature senescence as evidenced by the decrease in water content and total soluble protein content but those of soybean leaves was increased when stresses were recovered for 3days. The activities of APDX and GR subjected to drought or flooding were the decrease but those of enzymes were recovered when water stress was recovered. The activities of MDHAR with drought or flooding were on the decrease, whereas those of DHAR were increased, respectively. Antioxidant contents decreased continually subjected to drought or flooding but it recovered after 3 days subjected to water stresses.

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