• Title/Summary/Keyword: G proteins

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Proteome Analysis of Paenibacillus polymyxa E681 Affected by Barley

  • Seul, Keyung-Jo;Park, Seung-Hwan;Ryu, Choong-Min;Lee, Yong-Hyun;Ghim, Sa-Youl
    • Journal of Microbiology and Biotechnology
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    • v.17 no.6
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    • pp.934-944
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    • 2007
  • Paenibacillus polymyxa E681 is known to be able to suppress plant diseases by producing antimicrobial compounds and to promote plant growth by producing phytohormones, and secreting diverse degrading enzymes. In spite of these capabilities, little is known regarding the flow of information from the bacterial strain to the barley roots. In an attempt to determine the flow of information from the bacterial strain to barley roots, the strain was grown in the presence and absence of barley, and two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and MALDI-TOF mass spectrometry were used. 2D-PAGE detected approximately 1,000 spots in the cell and 1,100 spots in the supernatant at a pH 4-10 gradient. Interestingly, about 80 spots from each sample showed quantitative variations. Fifty-three spots from these were analyzed by MALDI-TOF mass spectrometry and 28 proteins were identified. Most of the cytosolic proteins expressed at higher levels were found in P. polymyxa E681 cells grown in the presence of barley rather than in the absence of barley. Proteins detected at a lower level in the surpernatant of P. polymyxa E68l cells grown in the presence of barley were lipoprotein, glucose-6-phosphate 1-dehydrogenase, heat-shock protein HtpG, spermidine synthase, OrfZ, ribonuclease PH, and coenzyme PQQ synthesis protein, and flagellar hook-associated protein 2 whereas proteins detected at a higher level in the surpernatant of P. polymyxa E681 cells grown in the presence of barley included D-alanyl-D-alanine ligase A, isopentenyl-diphosphate delta-isomerase, ABC transporter ATP-binding protein Uup, lipase. Many of the proteins belonging to plant-induced stimulons are associated with biosynthetic metabolism and metabolites of proteins and transport. Some of these proteins would be expected to be induced by environmental changes resulting from the accumulation of plant-secreted substances.

Directional adjacency-score function for protein fold recognition

  • Heo, Mu-Young;Cheon, Moo-Kyung;Kim, Suhk-Mann;Chung, Kwang-Hoon;Chang, Ik-Soo
    • Interdisciplinary Bio Central
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    • v.1 no.2
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    • pp.8.1-8.6
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    • 2009
  • Introduction: It is a challenge to design a protein score function which stabilizes the native structures of many proteins simultaneously. The coarse-grained description of proteins to construct the pairwise-contact score function usually ignores the backbone directionality of protein structures. We propose a new two-body score function which stabilizes all native states of 1,006 proteins simultaneously. This two-body score function differs from the usual pairwise-contact functions in that it considers two adjacent amino acids at two ends of each peptide bond with the backbone directionality from the N-terminal to the C-terminal. The score is a corresponding propensity for a directional alignment of two adjacent amino acids with their local environments. Results and Discussion: We show that the construction of a directional adjacency-score function was achieved using 1,006 training proteins with the sequence homology less than 30%, which include all representatives of different protein classes. After parameterizing the local environments of amino acids into 9 categories depending on three secondary structures and three kinds of hydrophobicity of amino acids, the 32,400 adjacency-scores of amino acids could be determined by the perceptron learning and the protein threading. These could stabilize simultaneously all native folds of 1,006 training proteins. When these parameters are tested on the new distinct 382 proteins with the sequence homology less than 90%, 371 (97.1%) proteins could recognize their native folds. We also showed using these parameters that the retro sequence of the SH3 domain, the B domain of Staphylococcal protein A, and the B1 domain of Streptococcal protein G could not be stabilized to fold, which agrees with the experimental evidence.

Label-free quantitative proteomic analysis of Panax ginseng leaves upon exposure to heat stress

  • Kim, So Wun;Gupta, Ravi;Min, Cheol Woo;Lee, Seo Hyun;Cheon, Ye Eun;Meng, Qing Feng;Jang, Jeong Woo;Hong, Chi Eun;Lee, Ji Yoon;Jo, Ick Hyun;Kim, Sun Tae
    • Journal of Ginseng Research
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    • v.43 no.1
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    • pp.143-153
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    • 2019
  • Background: Ginseng is one of the well-known medicinal plants, exhibiting diverse medicinal effects. Its roots possess anticancer and antiaging properties and are being used in the medical systems of East Asian countries. It is grown in low-light and low-temperature conditions, and its growth is strongly inhibited at temperatures above $25^{\circ}C$. However, the molecular responses of ginseng to heat stress are currently poorly understood, especially at the protein level. Methods: We used a shotgun proteomics approach to investigate the effect of heat stress on ginseng leaves. We monitored their photosynthetic efficiency to confirm physiological responses to a high-temperature stress. Results: The results showed a reduction in photosynthetic efficiency on heat treatment ($35^{\circ}C$) starting at 48 h. Label-free quantitative proteome analysis led to the identification of 3,332 proteins, of which 847 were differentially modulated in response to heat stress. The MapMan analysis showed that the proteins with increased abundance were mainly associated with antioxidant and translation-regulating activities, whereas the proteins related to the receptor and structural-binding activities exhibited decreased abundance. Several other proteins including chaperones, G-proteins, calcium-signaling proteins, transcription factors, and transfer/carrier proteins were specifically downregulated. Conclusion: These results increase our understanding of heat stress responses in the leaves of ginseng at the protein level, for the first time providing a resource for the scientific community.

Transduction of Familial Amyotrophic Lateral Sclerosis-related Mutant PEP-1-SOD Proteins into Neuronal Cells

  • An, Jae Jin;Lee, Yeom Pyo;Kim, So Young;Lee, Sun Hwa;Kim, Dae Won;Lee, Min Jung;Jeong, Min Seop;Jang, Sang Ho;Kang, Jung Hoon;Kwon, Hyeok Yil;Kang, Tae-Cheon;Won, Moo Ho;Cho, Sung-Woo;Kwon, Oh-Shin;Lee, Kil Soo;Park, Jinseu;Eum, Won Sik;Choi, Soo Young
    • Molecules and Cells
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    • v.25 no.1
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    • pp.55-63
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    • 2008
  • Amyotrophic lateral sclerosis (ALS) is a progressive neurodegenerative disorder characterized by the selective death of motor neurons. Mutations in the SOD1 gene are responsible for a familial form of ALS (FALS). Although many studies suggest that mutant SOD1 proteins are cytotoxic, the mechanism is not fully understood. To investigate the role of mutant SOD1 in FALS, human SOD1 genes were fused with a PEP-1 peptide in a bacterial expression vector to produce in-frame PEP-1-SOD fusion proteins (wild type and mutants). The expressed and purified PEP-1-SOD fusion proteins were efficiently transduced into neuronal cells. Neurones harboring the A4V, G93A, G85R, and D90A mutants of PEP-1-SOD were more vulnerable to oxidative stress induced by paraquat than those harboring wild-type proteins. Moreover, neurones harboring the mutant SOD proteins had lower heat shock protein (Hsp) expression levels than those harboring wild-type SOD. The effects of the transduced SOD1 fusion proteins may provide an explanation for the association of SOD1 with FALS, and Hsps could be candidate agents for the treatment of ALS.

A Study on the Improvement of Dietary Protein-efficiency by Supplement of the Panax Ginseng-by-products. (인삼의 부산물을 이용한 식의성 단백질의 효율 향상을 위한 연구)

  • 황우익;이성동
    • Journal of Ginseng Research
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    • v.3 no.1
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    • pp.1-34
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    • 1979
  • Our nation is confronted with the situation that the rice, a principal food, short of some essential amino acids, leads to imbalanced meals insufficient in the nutrient of Protein, to bring many difficulties in the elevation of nutritional state in our nation. While. our country has been produced much amounts of Panax Ginseng roots which has a stimulating effects on the metabolism of protein, lipid and nucleic acids in the body. And the leaf and trunk of Panax Ginseng were also produced a considerable amounts as the by-products. Author believe that these by-products (leaf and trunk) of Panax Ginseng might have some components possessing simillar activity with Panax Ginseng root although the quantity and qualify of the functional components may more or less be different. Therefore, this study was demised to observe the supplemental effect of the Panax Ginseng-by-Products on the dietary protein efficiency and nutritional state of rats. The feeds used for this experiment were rice containing 30% barely, fish four, and the leaf, trunk and small root of the Panax Ginseng, and the contents of the general nutrients including protein, lipid and carbohydrate etc. in each feed were analyzed for the combination of each feed. And, being based on analytical values of Protein in food. fish Pour as Protein source was added were rice containing 30% barely to be include 8.6 to 8.7%, 12%, 15% and 18% of protein. Then 2% of the leaf, trunk or small reef of Panax Ginseng was supplemented into each of above protein diet group, ton 16 kinds of diets were Prepared. The male albino rats from a Pure strain, weighing 70g to 80g. were used for experimental animals. They were maintained with coresponding fist for f and 8 weeks, and the growth rate, consumption of diets and protein, efficiency of feed and Protein in animals were determined. The lipids, proteins and cholesterols in serum and liver were also determined quantitatively after they were sacrificed in coresponding term. The results obtained are summarized as follows: 1. Body weigh of diet group containing 8.6 to 8.7%,12%, and 15% of protein are increased remarkably by supplement of 2% of the leaf or small root of Panax Ginseng in comparison with each of controls. But this tendency could not observed in diet group containing 18eA Proteins. 2. Feed efficiency showed same tendency in comparison with changes of gained body weight. Specially, in each of diets containing 8.7%, 12%, 15% and 18% of Proteins, supplement of the leaf of Panax Ginseng showed the better feed efficiency than supplement of the trunk or small root. 3. In feeding group for 8 weeks, protein efficiency showed worst efficiency in diet containing 18% proteins and showed the best efficiency was the diet group containing 12% Proteins. And the efficiency was improved according to supplement of the leaf of Panax Ginseng. 4. Nitrogen contents in serum and liver did not show large differences each other in all diet groups. But contexts of total cholesterol and 1ipid were decreased markedly in diet groups containing 12%, 15% and 18% of proteins in comparison with diet group containing 8.6% to 8.8% of proteins.

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Genetic effects of sterol regulatory element binding proteins and fatty acid-binding protein4 on the fatty acid composition of Korean cattle (Hanwoo)

  • Oh, Dong-Yep;Lee, Jea-Young;Jang, Ji-Eun;Lee, Seung-Uk
    • Asian-Australasian Journal of Animal Sciences
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    • v.30 no.2
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    • pp.160-166
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    • 2017
  • Objective: This study identifies single-nucleotide polymorphisms (SNP) or gene combinations that affect the flavor and quality of Korean cattle (Hanwoo) by using the SNP Harvester method. Methods: Four economic traits (oleic acid [C18:1], saturated fatty acids), monounsaturated fatty acids, and marbling score) were adjusted for environmental factors in order to focus solely on genetic effects. The SNP Harvester method was used to investigate gene combinations (two-way gene interactions) associated with these economic traits. Further, a multifactor dimensionality reduction method was used to identify superior genotypes in gene combinations. Results: Table 3 to 4 show the analysis results for differences between superior genotypes and others for selected major gene combinations using the multifactor dimensionality reduction method. Environmental factors were adjusted for in order to evaluate only the genetic effect. Table 5 shows the adjustment effect by comparing the accuracy before and after correction in two-way gene interactions. Conclusion: The g.3977-325 T>C and (g.2988 A>G, g.3977-325 T>C) combinations of fatty acid-binding protein4 were the superior gene, and the superior genotype combinations across all economic traits were the CC genotype at g.3977-325 T>C and the AACC, GACC, GGCC genotypes of (g.2988 A>G, g.3977-325 T>C).

The concentration of serum amyloid A and haptoglobin of pigs infected with Aujeszky′s disease virus (오제스키병에 감염된 돼지의 serum amyloid A와 haptoglobin의 농도 변화)

  • 오윤택;조정곤
    • Korean Journal of Veterinary Service
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    • v.25 no.1
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    • pp.45-52
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    • 2002
  • The acute phase serum protein response is a well-known general indicator of inflammation, trauma or other pathological conditions and its relevance for the monitoring of the health status of domestic animals is being increasingly realized. The changes in serum protein composition which occur after tissue damage represent a part of the systemic response of the injured animals which is mediated by pro-inflammatory cytokines such as TNF-$\alpha$, IL-6 and IL-1. These responses play a vital role in containing the tissue damage and enhancing the processes of repair and resolution. From a clinical perspective, the assay of acute phase proteins can provide a method for detecting inflammation. In animals, the most sensitive acute phase proteins are haptoglogin, serum amyloid A and at-acid glycoprotein in response to inflammatory condition. The aim of this study was to assess the diagnostic value of the concentrations of serum amyloid A(SAA) and haptoglobin(HP) in serum of pigs infected with Aujeszky's disease virus(ADV). Fifty pigs infected with ADV and 5 normal pigs were used in this experiment. The mean serum concentration of Shh of pigs infected with ADV was 96.8 $\pm$ 7.1 $\mu\textrm{g}$/㎖(range, 36.0∼187.5 $\mu\textrm{g}$/㎖) and that of normal pigs was 42.9$\pm$3.3 $\mu\textrm{g}$/㎖(range, 17.3∼127.8 $\mu\textrm{g}$/㎖). The mean serum concentration of HP of pigs infected with ADV was 1,164.4 $\pm$ 96.9 $\mu\textrm{g}$/㎖ (range, 790.2∼l,769.2 $\mu\textrm{g}$/㎖) and that of normal pigs was 675.4 $\pm$ 56.3 $\mu\textrm{g}$/㎖ (range, 650.0-690.4 $\mu\textrm{g}$/㎖). The mean concentrations of SAA and HP in serum of pigs infected with ADV compared with those of normal pigs showed approximately a two-fold. It was concluded that the concentrations of Shh and HP in serum may proved to be diagnostic marker of Aujeszky's disease.

A Comparison of Two Methods for the Extraction of Lactoferrin-binding Proteins from Streptococcus uberis (Streptococcus uberis의 락토페린 결합단백질 추출을 위한 두 가지 방법의 비교)

  • Park, Hee-Myung;Yoo, Jong-Hyun;Almeida, Raul A.;Oliver, Stephen P.
    • Journal of Veterinary Clinics
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    • v.24 no.3
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    • pp.305-307
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    • 2007
  • Lactoferrin-binding proteins (LBP) has not been well characterized in Streptococcus uberis isolated from milk of bovine mastitis and to date this protein is considered to be an important virulence factor in Streptococcal mastitis. To determine the more efficient extraction method of LBP from four S. uberis strains, we used two different extraction methods (mutanolysin and sodium dodecyl sulfate) in this study. Bacterial proteins extracted were electrophoresed by 10% polyacrylamide gels in the presence of sodium deodecyl sulfate and gels were transferred onto nitrocellulose membrane. Rabbit anti-bovine lactoferrin antibody and HRP-conjugated donkey anti-rabbit IgG antibody were used to detect LBP. This Western blotting analysis demonstrates that extraction method with SDS extracted 110 kDa and 112 kDa LBPs more efficiently compared to the mutanolysin extraction method.

Abscisic Acid Binding to Extracts from Normal and Viviparous-1 Mutant Aleurone Layers of Zea mays L.

  • Bai, Dong-Gyu
    • Journal of Plant Biology
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    • v.37 no.2
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    • pp.151-158
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    • 1994
  • Aleurone layers of normal and vp1 mutant maize kernels were extracted and centrifuged at 100,000g to yield a cytosol fraction. Binding of [3H]ABA cis, trans (+)ABA to a soluble macromolecular components present in the cytosol was demonstrated by Sephadex chromatography and non-denaturing PAGE. The binding component was of high molecular weight and seems to be an aggregate of proteins. A rapid DEAE-cellulose filter method for assaying bound [3H]ABA to a soluble protein was adapted. Binding assays were performed with cytosol that had been preheated or incubated with several enzymes, indicating that heat and protease treatments disrupted the binding. This suggested that binding occurred to proteins. Some properties of the ABA binding proteins were described. The [3H]ABA binding were reduced dramatically when unlabeled ABA was added as a competitor, suggesting a specific binding of [3H]ABA. Gel filtration profiles and autoradiogram of [3H]ABA binding showed no difference in the binding components of Vp1 and vp1/vp1 mutant cytosol, indicating that Vp1 protein is not a sole ABA binding protein.

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