• 제목/요약/키워드: 2a protein

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Effects of Plasma on the Surface of Protein Chip Plates (단백질 칩 기판의 플라즈마 효과)

  • Hyun, J.W.;Kim, N.Y.
    • Journal of the Korean Vacuum Society
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    • 제17권6호
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    • pp.549-554
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    • 2008
  • Nickel Chloride coated protein chip plates were developed by using a spin coating method after $H_2$ plasma treatment. The adsorption ability of histidine tagged protein was investigated at various times of plasma treatment. The properties of the nickel chloride and protein on the surface of the slides were assayed using particle size analysis and the extent of the protein adsorption was determined by using a bio imaging analyzer system. The results show that the ability of protein adsorption decreased as increasing the time of $H_2$ plasma treatment. The mechanism on the ability of protein adsorption at the plate surface is discussed on results and discussions. The results also suggest that the surface stabilization of protein chip plates treated by plasma technology may be applicable in biosensor markets.

Soluble expression and purification of synthetic human bone morphogenetic protein-2 in Escherichia coli

  • Ihm, Hyo-Jin;Yang, Seung-Ju;Huh, Jae-Wan;Choi, Soo-Young;Cho, Sung-Woo
    • BMB Reports
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    • 제41권5호
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    • pp.404-407
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    • 2008
  • A 345-bp gene that encodes human bone morphogenetic protein-2 (hBMP-2) has been synthesized. The codon usage of the resulting gene was modified to include those triplets that are utilized in highly expressed Escherichia coli genes. The hBMP-2 gene was efficiently expressed in E. coli as a soluble and active protein. Since the recombinant hBMP-2 was readily solublized, no further solublization steps were required throughout purification. No additional tagging residues were introduced into the synthetic hBMP-2 gene product. The developed synthetic gene is a promising approach for scaling-up the soluble expression of hBMP-2.

A WD40 Repeat Protein, Arabidopsis Sec13 Homolog 1, May Play a Role in Vacuolar Trafficking by Controlling the Membrane Association of AtDRP2A

  • Lee, Myoung Hui;Lee, Sung Hoon;Kim, Heyran;Jin, Jing Bo;Kim, Dae Heon;Hwang, Inhwan
    • Molecules and Cells
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    • 제22권2호
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    • pp.210-219
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    • 2006
  • Dynamin-related protein 2A (AtDRP2A, formally ADL6), a member of the dynamin family, is critical for protein trafficking from the TGN to the central vacuole. However, the mechanism controlling its activity is not well understood in plant cells. We isolated Arabidopsis sec13 homolog1 (AtSeh1) that interacts with AtDRP2A by a yeast two-hybrid screening. AtSeh1 has four WD40 motifs and amino acid sequence homology to Sec13, a component of COPII vesicles. Coimmunoprecipitation and protein pull-down experiments demonstrated specific interaction between AtSeh1 and AtDRP2A. AtSeh1 bound to the pleckstrin homology domain of AtDRP2A in competition with the C-terminal domain of the latter, and this resulted in inhibition of the interaction between AtDRP2A and PtdIns3P in vitro. AtSeh1 localized to multiple locations: the nucleus, the prevacuolar compartment and the Golgi complex. Based on these results we propose that AtSeh1 plays a role in regulating cycling of AtDRP2A between membrane-bound and soluble forms.

Molecular and Biochemical Studies on the DNA Replication of Bacteriophage T7: Functional Analysis of Amino-terminal Region of Gene 2.5 Protein

  • Kim, Young-Tae;Lee, Sung-Gu;Kim, Hak-Jun
    • BMB Reports
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    • 제28권6호
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    • pp.484-489
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    • 1995
  • The product of bacteriophage T7 gene 2.5 is a single-stranded DNA binding protein and plays an important role in T7 DNA replication, recombination, and repair. Genetic analysis of T7 phage defective in gene 2.5 shows that the gene 2.5 protein is essential for T7 DNA replication and growth (Kim and Richardson, 1993). The C-terminal truncated gene 2.5 protein ($GP2.5-{\Delta}21C$) cannot substitute for wild-type gene 2.5 protein in vivo; suggesting that the C-terminal domain of gene 2.5 protein is essential for protein-protein interactions (Kim and Richardson, 1994; J. Biol. Chem. 269, 5070-5078). Truncated gene 2.5 proteins lacking 19 residues ($GP2.5-{\Delta}19N$) and 39 residues ($GP2.5-{\Delta}39N$) from the amino-terminal domain were constructed by in vitro mutagenesis. $GP2.5-{\Delta}19N$ can support the growth of T7 phage lacking gene 2.5 while $GP2.5-{\Delta}39N$ cannot substitute for wild-type gene 2.5 protein in vivo; however, its ability to bind to single-stranded DNA is not affected. These results clearly demonstrate that the 20~39 amino-terminal region of gene 2.5 protein is required for T7 growth in vivo but may not be involved in DNA binding activity.

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Optimum Dietary Protein Level of Ayu (Plecoglossus altivelis) (은어 사료의 적정 단백질 함량)

  • 이상민;김경덕
    • Journal of Aquaculture
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    • 제12권2호
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    • pp.145-153
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    • 1999
  • This study was conducted to determine the protein requirement of ayu (Plecoglossus altivelis). Two replicate groups of fish initially averaging 6.6 g were fed the five isocaloric diets containing different protein level from 29% to 57% in a flow-through freshwater system for 25 days. White fish meal was used as a sole protein source. Weight gain and feed efficiency of fish increased significantly with dietary protein level up to 43% (P<0.05) with no additional response above this level. Protein and lipid retention, moisture, protein and lipid contents of body were not affected by dietary protein levels (P>0.05). Daily protein intake increased significantly with dietary protein level, whereas protein efficiency ratio of fish fed the 57% dietary protein decreased (P<0.05). The data obtained in this study indicate that a 43% dietary protein level could be recommended for the optimum growth of ayu.

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The Soluble Expression of the Human Renin Binding Protein Using Fusion Partners: A Comparison of ubquitin, Thioredoxin, Maltose Binding Protein-and NusA

  • Lee, Chung;Lee, Sun-Gu;Saori Takahashi;Kim, Byung-Gee
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권2호
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    • pp.89-93
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    • 2003
  • human renin binding protein (hRnBp), showing N-acetylglucosamine-2-epimerase activity, was over-expressed in E. coli, but was mainly present as an inclusion body. To improve its solubility and activity, ubiquitin (Ub), thioredoxin (Trx), maltose binding protein (MBP) and NusA, were used as fusion partners. The comparative solubilities of the fusion proteins were, from most to least soluble: NusA, MBP, Trx, Ub. Only the MBP fusion did not significantly reduce the activity of hRnBp, but enhanced the stability. The Origami (DE3), permitting a more oxidative environment for the cytoplasm in E. coli; helped to increase its functional activity.

The Effects of Chronic Carbamazepine Administration on Protein Kinase A and Protein Kinase C Activities in Rat Brain (카바마제핀 장기 투여가 백서(白鼠) 뇌의 Protein Kinase A와 Protein Kinase C 활성도에 미치는 영향)

  • Rheem, Doo-Won;Kim, Leen;Suh, Kwang-Yoon
    • Korean Journal of Biological Psychiatry
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    • 제5권2호
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    • pp.227-234
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    • 1998
  • Objective : Many evidences suggest that patients with bipolar disorder have functional abnormalities in their postreceptor signal transduction pathways, and mood stabilizing effect of lithium is exerted by modulating this dysfunctioning system. Carbamazepine, an antiepileptic agent, is also known to be effective in the treatment and prevention of bipolar disorder. But the precise mechanism of action of the drug is still poorly understood. This study was performed to elucidate the possible therapeutic mechanism of carbamazepine. Method : The effects of chronic carbamazepine administration on protein kinase A and protein kinase C activities in frontal cortex of rat brain after 2 weeks of drug administration were measured and compared with those of control subjects. Results : Mean(${\pm}SE$) value of activity(phosphate transfer ${\mu}mol/mg$ of $protein{\cdot}min$) of protein kinase A in control and test group was $0.249563{\pm}0.036$ and $0.539853{\pm}0.078$, and that of protein kinase C was $0.654817{\pm}0.053$ and $1.146205{\pm}0.052$ respectively, being increased in test group. And differences between the two groups were statistically significant for both enzymes(protein kinase A ; p<0.01, protein kinase C ; p<0.001). Conclusion : These results show that chronic carbamazepine administration increases protein kinase A and C activities, and concerning the possible mode of therapeutic action in bipolar disorder it is suggested that enhanced enzymes phosphorylate receptor-G-protein-effector complexes to dampen hyperfunctioning neuronal activity and thus stabilize the system.

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Effect of Lowering Dietary Protein with Constant Energy to Protein Ratio on Growth, Body Composition and Nutrient Utilization of Broiler Chicks

  • Kamran, Z.;Sarwar, M.;Nisa, M.;Nadeem, M.A.;Ahmad, S.;Mushtaq, T.;Ahmad, T.;Shahzad, M.A.
    • Asian-Australasian Journal of Animal Sciences
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    • 제21권11호
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    • pp.1629-1634
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    • 2008
  • A trial was conducted to determine the effect of low crude protein (CP) diets with constant metabolizable energy to crude protein (ME:CP) ratio on growth, body composition and nutrient utilization of broiler chicks from 1 to 26 days of age. Four dietary treatments having four levels of CP and ME as 23, 22, 21 and 20% and 3,036, 2,904, 2,772 and 2,640 kcal/kg, respectively, were formulated and a ME:CP ratio of 132 was maintained in all the diets. Digestible lysine was maintained at 1.10 of the diet. A total of 1,760 day-old Hubbard broiler chicks were randomly divided into 16 experimental units and each diet was offered to four experimental units at random. Feed intake was increased (p<0.05) while weight gain and feed conversion ratio were adversely affected (p<0.05) when the diets with low CP and ME were fed to broilers. Total protein intake and total ME intake were linearly decreased (p<0.05) and protein efficiency ratio and energy efficiency ratio were lower (p<0.05) than in the chicks fed dietary regimen with 22% CP and 2,904 kcal/kg ME. The whole body analysis of the birds revealed that chicks fed the lowest dietary regimens retained less (p<0.05) nitrogen and more ether extract than chicks fed the control diet, however, body dry matter, total body ash and fat free body protein were not affected. Similarly, protein and energy utilization were also unaffected by the dietary treatments. In summary, chicks fed low CP diets with constant ME:CP ratio grew slower, used feed less efficiently and retained less protein and more body fat than chicks fed the control diet.

Effects of Dietary Blood Meal as a Protein Source in Growing Common Carp (Cyprinus carpio) (성장기 잉어(Cyprinus carpio) 사료에 있어서 단백질원으로서의 혈분 첨가효과)

  • Song Min-Heon;Lee Kyeong-Jun;Bai Sungchul
    • Journal of Aquaculture
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    • 제8권4호
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    • pp.343-354
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    • 1995
  • A series of feeding experiments were conducted to study the possible utilization of blood meal as a dietary protein source in growing common carp, Cyprinus carpio. Diets were formulated on isonitrogenous and isocaloric basis of $40\%$ crude protein and 3,640 kcal/kg diet : diet 1 (100 FMP, control), $100\%$ fish meal Protein (FMP) : diet 2 (25 BMP), $75\%$ $FMP+25\%$ blood meal protein (BMP) : diet 3 (50 BMP), $50\%$ $FMP+50\%$ BMP : diet 4 (75 BMP), $25\%$ $FMP+75\%$ BMP : d;et 5 (100 BMP), $100\%$ BMP. As the dietary protein sources, $34.2\%$ of animal protein were supplied by FMP and/or BMP, and approximately $65.8\%$ of plant protein were used. In the first experiment, weight gain and feed efficiency were improved with increased level of blood meal protein in the diets. Weight gain and feed efficiency from fish fed diets 4 and S were significantly higher (P<0.05) than those from fish fed diets 1, 2 and 3. The second experiment was designed as a cross-over study to prove the first experiment's results. This cross-over study shows that weight gain from fish fed diet 5 is greater than that from fish fed diet 1. The third experiment was conducted to compare palatability between diet 1 (100 FMP) and 5 (100 BMP). The data from this palatability study indicated that the palatability of diet 5 was lower than that of diet 1 initially, however, the palatability of iiet 5 was improved and not worse than that of diet 1 within a week. Therefore, these findings may suggest that the fish meal can be replaced with blood meal completely in growing common carp diets.

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Effect of NaCl, Phosphate and pH on the Functional Properties of a Mixed System of Pork Myofibrillar and Plasma Proteins (소금, 인산염, pH가 돼지 혈장단백질과 근원섬유단백질 혼합물의 기능적 특성에 미치는 영향)

  • Kim, Cheon-Jei;Han, Eui-Su
    • Korean Journal of Food Science and Technology
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    • 제23권4호
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    • pp.428-432
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    • 1991
  • This study was carried out to investigate the effect of NaCl, pH and phosphate on the functional properties of a mixed system of plasma protein and myofibrillar proteins. The solubility of plasma protein, myofibrillar protein and the mixture (plasma+myofibrillar protein) increased according to the increase of NaCl concentration ($0{\sim}4%$) and pH $pH4{\sim}8$). The solubility, emulsifying activity and capacity of the mixture were lower than those of plasma protein, whereas higher than those of myofibrillar protein. The gel strength of the mixture and myofibrillar protein showed a significant increase when NaCl concentration was increased from 2 to 3%. The gel strength of myofibrillar protein increased about four times when 0.3% polyphosphate added to the sample containing 2% NaCl, whereas the moisture loss of the mixture and myofibrillar protein decreased significantly. The gel strength of plasma protein, myofibrillar protein and the mixture increased slightly at $3{\sim}5%$ protein concentration, whereas the gel strength of those increased significantly as the protein concentration increased from 5 to 9%.

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