• Title/Summary/Keyword: P1 protein

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Purification and Characterization of a Novel Antifungal Protein from Paenibacillus macerans PM1 Antagonistic to Rice Blast Fungus, Pyricularia oryzae

  • Bae, Dong-Won;Kawk, Weon-Sik;Lee, Joon-Taek;Son, Dae-Young;Chun, Sung-Sik;Kim, Hee-Kyu
    • Journal of Microbiology and Biotechnology
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    • v.10 no.6
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    • pp.805-810
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    • 2000
  • An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to $100^{\circ}C$. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein ($5.9{\mu} g/ml$) was limited to $9{\pm}3.2%$ only, compared with $69{\pm}2.4%$ of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein ($5.9{\mu} g/ml$)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.

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Functional Implications in Apoptosis by Interferon Inducible Gene Product 1-8D, the Binding Protein to Adenovirus Preterminal Protein

  • Joung, In-Sil;Angeletti, Peter C.;Engler, Jeffrey A.
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.295-299
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    • 2003
  • Adenovirus (Ad) precursor to the terminal protein (pTP) plays an essential roles in the viral DNA replication. Ad pTP serves as a primer for the synthesis of a new DNA strand during the initiation step of replication. In addition, Ad pTP forms organized spherical replication foci on the nuclear matrix (NM) and anchors the viral genome to the NM. Here we identified the interferon inducible gene product 1-8D (Inid) as a pTP binding protein by using a two-hybrid screen of a HeLa cDNA library. Of the clones obtained in this assay, nine were identical to the Inid, a 13-kDa polypeptide that shares homology with genes 1-8U and Leu-13/9-27, most of which have little known functions. The entire open reading frame (ORF) of Inid was cloned into the tetracycline inducible expression vector in order to determine the biological functions related with adenoviral infection. When Inid was introduced to the cells along with adenoviruses, fifty to sixty percent of Ad-infected cells expressing Inid had rounded morphology, which was suggestive of apoptosis. Results from the terminal deoxynucleotidyl transferase (TdT) and DNA fragmentation assays confirmed that Inid induces apoptosis in Ad-infected or in uninfected cells. The Inid binding to pTP may target the cell for apoptotic destruction as a host defense mechanism against the viral infection.

Maternal Low-protein Diet Alters Ovarian Expression of Folliculogenic and Steroidogenic Genes and Their Regulatory MicroRNAs in Neonatal Piglets

  • Sui, Shiyan;Jia, Yimin;He, Bin;Li, Runsheng;Li, Xian;Cai, Demin;Song, Haogang;Zhang, Rongkui;Zhao, Ruqian
    • Asian-Australasian Journal of Animal Sciences
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    • v.27 no.12
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    • pp.1695-1704
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    • 2014
  • Maternal malnutrition during pregnancy may give rise to female offspring with disrupted ovary functions in adult age. Neonatal ovary development predisposes adult ovary function, yet the effect of maternal nutrition on the neonatal ovary has not been described. Therefore, here we show the impact of maternal protein restriction on the expression of folliculogenic and steroidogenic genes, their regulatory microRNAs and promoter DNA methylation in the ovary of neonatal piglets. Sows were fed either standard-protein (SP, 15% crude protein) or low-protein (LP, 7.5% crude protein) diets throughout gestation. Female piglets born to LP sows showed significantly decreased ovary weight relative to body weight (p<0.05) at birth, which was accompanied with an increased serum estradiol level (p<0.05). The LP piglets demonstrated higher ratio of bcl-2 associated X protein/B cell lymphoma/leukemia-2 mRNA (p<0.01), which was associated with up-regulated mRNA expression of bone morphogenic protein 4 (BMP4) (p<0.05) and proliferating cell nuclear antigen (PCNA) (p<0.05). The steroidogenic gene, cytochrome P450 aromatase (CYP19A1) was significantly down-regulated (p<0.05) in LP piglets. The alterations in ovarian gene expression were associated with a significant down-regulation of follicle-stimulating hormone receptor mRNA expression (p<0.05) in LP piglets. Moreover, three microRNAs, including miR-423-5p targeting both CYP19A1 and PCNA, miR-378 targeting CYP19A1 and miR-210 targeting BMP4, were significantly down-regulated (p<0.05) in the ovary of LP piglets. These results suggest that microRNAs are involved in mediating the effect of maternal protein restriction on ovarian function through regulating the expression of folliculogenic and steroidogenic genes in newborn piglets.

Structural stability of CD1 domain of human mitotic checkpoint serine/threonine-protein kinase, Bub1

  • Kim, Hyun-Hwi;Song, Hyun-Kyu;Lee, Bong-Jin;Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.2
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    • pp.88-94
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    • 2015
  • Bub1 is one of the spindle checkpoint proteins and plays a role in recruitment of the related proteins to kinetochore. Here, we studied the structural characteristic of the evolutionarily conserved 160 amino acid region in the N-terminus (hBub1 CD1), using Circular Dichroism (CD) and NMR. Our CD results showed that hBub1 CD1 is a highly helical protein and its structure was affected by pH: as pH was elevated to basic pH, the helical propensity increased. This could be related to the surface charge of the hBub1 CD1. However, the structural change did not largely depend on the salt concentration, though the thermal stability a little increased. The previous NMR analysis revealed that the hBub1 CD1 adopts eight helices, which is consistent with the CD result. Our result would be helpful for evaluating the molecular mechanism of the hBub1 CD1 and protein-protein interactions.

Emulsion Properties of Small Red Bean Protein Isolates (분리 팥 단백질의 유화특성에 관한 연구)

  • Kim, Hyon-Jung;Sohn, Kyung-Hee;Park, Hyun-Kyung
    • Korean journal of food and cookery science
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    • v.6 no.4 s.13
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    • pp.9-14
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    • 1990
  • The emlsifying properties of small red bean protein isoates were evaluated through their emulsion capacity and stability of the resulting emulsions. The influence of pH, Sodium Chloride and heat treatment on the efficiency of small red bean protein isolates as emulsifying agents was studied. The surface hydrophobicity (So) of small red bean protein islates also examined. The results were obtained as follows; 1. The emusion capacity of small red bean protein isolates was high at pH 11, low at pH 3 and decreased by heat treament. With addition of NaCl, emulsion capacity decreased steadily and showed lowest value when 0.2M NaCl was added. 2. The emulsion stability at pH 4.5 and heat treatment over $60^{\circ}C$ decreased emulsion stability at pH 4.5. When NaCl was added, emulsion stability was generally increased. 3. The surface hydrophobicity of small red bean protein isolates showed the highest value at pH 3 and lowest at pH 11 and increased as the heating temperature increased When 0.2 M NaCl was added, surface hydrophobicity also increased at pH 4.5.

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Effects of Non-protein Energy Intake on Whole Body Protein Synthesis, Nitrogen Retention and Glucose Turnover in Goats

  • Fujita, Tadahisa;Kajita, Masahiro;Sano, Hiroaki
    • Asian-Australasian Journal of Animal Sciences
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    • v.20 no.4
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    • pp.536-542
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    • 2007
  • The responses of whole body protein and glucose kinetics and of nitrogen (N) metabolism to non-protein energy intake (NPEI) were determined using an isotope dilution approach and measurement of N balance in three adult male goats. The diets containing 1.0, 1.5 and 2.0 times ME maintenance requirement, with fixed intake of CP (1.5 times maintenance) and percentage of hay (33%), were fed twice daily for each 21 d experimental period. After an adaptation period of 11 d, N balance was determined over 3 d. On day 17, whole body protein synthesis (WBPS) and glucose irreversible loss rate (ILR) were determined during the absorptive state by a primed-continuous infusion of [$^2H_5$]phenylalanine, [$^2H_2$]tyrosine, [$^2H_4$]tyrosine and [$^{13}C_6$]glucose, with simultaneous measurements of plasma concentrations of metabolites and insulin. Ruminal characteristics were also measured at 6 h after feeding over 3 d. Nitrogen retention tended to increase (p<0.10) with increasing NPEI, although digestible N decreased linearly (p<0.05). Increasing NPEI decreased (p<0.01) ammonia N concentration, but increased acetate (p<0.05) and propionate (p<0.05) concentrations in the rumen. Despite decreased plasma urea N concentration (p<0.01), increased plasma tyrosine concentration (p<0.05), and trends toward increased plasma total amino N (p<0.10) and phenylalanine concentrations (p<0.10) were found in response to increasing NPEI. Increasing NPEI increased ILR of both glucose (p<0.01) and phenylalanine (p<0.05), but did not affect ($p{\geq}0.10$) that of tyrosine. Whole body protein synthesis increased (p<0.05) in response to increasing NPEI, resulting from increased utilization rate for protein synthesis (p<0.05) and unchanged hydroxylation rate of phenylalanine ($p{\geq}0.10$). These results suggest that increasing NPEI may enhance WBPS and glucose turnover at the absorptive state and improve the efficiency of digestible N retention in goats, with possibly decreased ammonia and increased amino acid absorption. In addition, simultaneous increases in WBPS and glucose ILR suggest stimulatory effect of glucose availability on WBPS, especially when sufficient amino acid is supplied.

Very low protein diet plus ketoacid analogs of essential amino acids supplement to retard chronic kidney disease progression

  • Satirapoj, Bancha;Vongwattana, Peerapong;Supasyndh, Ouppatham
    • Kidney Research and Clinical Practice
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    • v.37 no.4
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    • pp.384-392
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    • 2018
  • Background: A very low protein diet (VLPD) with ketoacid analogs of essential amino acids (KA/EAA) administration can remarkably influence protein synthesis and metabolic disturbances of patients with advanced chronic kidney disease (CKD), and may also slow the decline in renal function. Methods: A retrospective cohort study was carried out to monitor renal progression and metabolic and nutritional status among 140 patients with CKD stage III or IV. One group (n = 70) was on a low protein diet (LPD) with 0.6 g of protein intake, and another group (n = 70) was on a VLPD with 0.3 g of protein and KA/EAA supplementation of 100 mg/kg/day for 12 months. Results: At 12-month follow-up, estimated glomerular filtration rate (GFR) significantly decreased from $41.6{\pm}10.2$ to $36.4{\pm}8.8mL/min/1.73m^2$ (P < 0.001) and urine protein increased from $0.6{\pm}0.5$ to $0.9{\pm}1.1g/day$ (P = 0.017) in the LPD group, but no significant changes in estimated GFR and urine protein were found in the VLPD plus KA/EAA group. A significant mean difference in rate of change in estimated GFR ($-5.2{\pm}3.6mL/min/1.73m^2$ per year; P < 0.001) was observed between the two groups. After Cox regression analysis, treatment with VLPD plus KA/EAA significantly protected against the incidence of declining GFR > 10% annually (adjusted hazard ratio, 0.42; 95% confidence interval, 0.23-0.79; P = 0.006) and significant correlations were found between using VLPD plus KA/EEA and increased GFR. Conclusion: VLPD supplementation with KA/EAA is associated with delayed renal progression while preserving the nutritional status in the patients with CKD. Co-administration of VLPD and KA/EAA may prove an effective alternative to conservative management of CKD.

Extraction and Purification of Rapeseed Protein (유채박 단백질의 추출 및 정제에 관한 연구)

  • Lee, Jang-Soon;Kang, Dong-Sub;Kang, Yeung-Joo
    • Korean Journal of Food Science and Technology
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    • v.22 no.7
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    • pp.780-785
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    • 1990
  • In order to establish the effective extraction and purification process of rapeseed protein, the extraction solvents were compared with one another ; and the residues of glucosinolate and phytate and the extraction yield of protein, which had been extracted by 1% sodium hexa mata-phosphate(SHMP) and purified through isoelectric precipitation, acid-washing and UF concentration, were investigated. As for the condition for extraction of rapeseed proteins, the solvent of 1% SHMP(pH 8.0) turned out the most appropriate ; so far as the purification process for the elimination of glucosinolate and phytate was concerned, the acid-washing twice or the process of the acid-washing once and UF concentration was considered the most effective. The yield and content of rapeseed protein were 37.1% and 75.3% respectively in the case of the acid-washing twice, 42.1% and 72.4% respectively in the case of the acid-washing once and UF concentraction, Consequently, with the elimination effects of glucosinolate and phytate put into consideration, the process of isoelectric precipitation, acid-washing once(pH 3.5), neutralizing(pH 7.5), UF concentration and then freeze drying proved the most effective purification process.

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Drug-Biomacromolecule Interaction (III) 1-Anilinonaphthalene-8-sulfonate Binding to Bovine Serum Albumin by Fluorescence (약물과 생체 고분자간의 상호작용(III) 형광측정법에 의한 1-Anilinonaphthalene-8-sulfonate와 Bovine Serum Albumin간의 결합에 관한 연구)

  • 김종국;안해영;양지선;김양배;유병설
    • YAKHAK HOEJI
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    • v.26 no.2
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    • pp.85-90
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    • 1982
  • The binding of the 1-anilinonaphthalene-8-sulfonate(ANS) to bovine serum albumin was studied by fluorescence spectroscopy. The effect of pH, ionic strength, and protein concentration on the binding of ANS to protein were compared. The binding between ANS and protein was dependent on pH and ionic strength. It seems that both hydrophobic binding and some electrostatic forces are involved in the binding of ANS to protein. The binding constants for ANS increased with increasing protein concentration. This suggests the possibility of a sharing of one ANS molecule by more than one protein molecule at relatively high protein concentration.

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Anti-inflammatory Effect of Cornus Officinalis fruit extract and Cornus Officinalis Fruit Cheonghyeol Plus in Human Umbilical Vein Endothelial Cell (인간 제대정맥 내피세포에서 산수유와 산수유청혈플러스의 항염증효과)

  • Jeong-hui Kim;Ho-ryong Yoo;In-chan Seol;Yoon-sik Kim
    • The Journal of Korean Medicine
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    • v.43 no.3
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    • pp.106-121
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    • 2022
  • Objectives: The purpose of this study was to investigate the anti-inflammatory effect of Cornus Officinalis fruit extract(CE) and Cornus Officinalis Fruit Cheonghyeol Plus(CCP) in Human Umbilical Vein Endothelial Cell. Methods: We measured cell viability of CE, CCP and treated HUVEC with TNF-α. We measured the mRNA expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, the protein expression levels of KLF2, eNOS, MCP-1, ICAM-1, VCAM-1, and the protein phosphorylation level of ERK, JNK, p38 and the biomarker expression levels of MCP-1, ICAM-1, VCAM-1. Results: 1.CE incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ compared to the control group. CE decresed the mRNA, protein and biomarker expression levels of MCP-1,ICAM-1,VCAM-1 at concentrations of 100㎍/㎖ compared to the control group. CE decresed the protein phosphorylation level of p38 at concentrations of 100㎍/㎖ compared to the control group. 2. CCP incresed the mRNA, protein expression levels of KLF2, eNOS at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the mRNA, protein and biomarker expression levels of MCP-1, ICAM-1, VCAM-1 at concentrations of 100㎍/㎖ or more compared to the control group. CCP decresed the protein phosphorylation level of ERK at concentrations of 100㎍/㎖ or more, p38 at concentrations of 200㎍/㎖ or more, and JNK at concentrations of 400㎍/㎖ compared to the control group. Conclusions: These results present that CE and CCP has anti-inflammatory effect in HUVEC. So, it could help treat or prevent inflammation in vein caused by dyslipidemia and contribute prevention of cardiovascular and cerebrovascular cerebrovascular diseases.