• Title/Summary/Keyword: anionic protein

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pH-Sensitive Dynamic Swelling Behavior of Glucose-containing Anionic Hydrogels (글루코스를 함유한 음이온 하이드로젤의 pH 감응성 동적 팽윤거동)

  • Kim, Bumsang
    • Korean Chemical Engineering Research
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    • v.43 no.2
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    • pp.299-304
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    • 2005
  • There have been many efforts to use anionic hydrogels as oral protein delivery carriers due to their pH-responsive swelling behavior. The dynamic swelling behavior of poly(methacrylic acid-co-methacryloxyethyl glucoside) [P(MAA-co-MEG)] hydrogels was investigated to determine the mechanism of water transport through these anionic hydrogels. The exponential relation $M_t/M_{\infty}=kt^n$ was used to calculate the exponent, n, describing the Fickian or non-Fickian behavior of swelling polymer networks. The mechanism of water transport through these gels was significantly affected by the pH of the swelling medium. The mechanism of water transport became more relaxation-controlled in the swelling medium of pH 7.0 that was higher than the $pK_a$ of the gels. Experimental results of time-dependent swelling behavior of the gels were analyzed with several mathematical models. Using ATR-FTIR spectroscopy, the effect of ionization of the carboxylic acid groups in the polymer networks on the water transport mechanism was investigated.

Physiological Relevance of Salt Environment for in vitro recA System

  • Kim, Jong-Il
    • Journal of Microbiology
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    • v.37 no.2
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    • pp.59-65
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    • 1999
  • RecA protein can promote strand assimilation, homologous pairing, and strand exchange. All these reactions require DNA-dependent ATP hydrolysis by recA protein, and the activities of recA protein are affected by the ionic environment. In this experiment, DNA-dependent ATPase activity showed different sensitivity to anionic species. ATP hydrolysis and strand exchange were relatively sensitive to salt in the reactions with NaCl, strongly inhibited at 100 mM NaCl. However, the inhibition by sodium acetate or sodium glutamate was not observed at 50∼100 mM concentration. Addition of sodium glutamate to the standard reaction condition increased the apparent efficiency of ATP hydrolysis during strand exchange. The condition including 50∼100 mM sodium-glutamate might be similar to the physiological condition.

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Membrane Insertion of Cytochrome P450 1A2 Promoted by Anionic Phospholipids

  • Yun, Chul-Ho
    • Proceedings of the Korean Biophysical Society Conference
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    • 1998.06a
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    • pp.16-16
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    • 1998
  • The role of phospholipids in the membrane binding and subsequent insertion of the microsomal protein rabbit cytochrome P450 (P450) lA2 into phospholipid bilayers was investigated. The insertion of P450 lA2 into phospholipid bilayers was determined by the amount of quenching of Trp fluorescence of P450 lA2 by pyrene and brominated and doxyl-labeled phospholipids.(omitted)

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Isolation and Purification of Lectin from Phaseolus radiatus (녹두로부터 렉틴 성분의 분리 정제)

  • 정시련;홍승수;전경희
    • YAKHAK HOEJI
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    • v.27 no.3
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    • pp.221-227
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    • 1983
  • New lectins, lymphoagglutinating lectins from mung beans (MBLA) have been isolated and purified. Mung beans crude extracts were made with 0.15M NaCl and these were purified through anionic exchange chromatography. Four fractions were obtained from DEAE Sephadex A-50 by salt gradients elution. Lectin activity, enzyme activity, protein assay, identification of purity by polyacrylamide gel electrphoresis and immunochemical studies were carried out with these four fractions. Through these results, it can be suggested that 0.2M fraction is newly found potent MBLA. There were some relationships with MBLA and L-PHA but no similarities were observed between MBLA and E-PHA.

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Measurement of Molecular Weight and Heating Properties of Korean White Ginseng Protein (백삼 단백질의 가열특성과 분자량 측정)

  • 박상욱
    • The Korean Journal of Food And Nutrition
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    • v.17 no.1
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    • pp.66-71
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    • 2004
  • This study was carried out to elucidate the molecular weight and the heating properties of Korean white ginseng protein by CM-cellulose column chromatography and electrophoresis. Thermostable protein contents were 0.17% in xylem-pith and 0.15% in cortex-epidermis of tap root by 90min of heating. The contents of thermostable protein were decrease after 90min of heating. By Electrophoresis, seven bands of 66, 45, 29, 24, 22, 20, 12kD were observed up to 30min of heating, but the band of 22kD was disappeared after 60min. of heating. The cationic protein content of thermostable protein fraction (28.24%) was higher than the anion protein content(0.80%). The molecular weight of thermostable protein fractions were 66kD, 55kD, 36kD and those of thermolabile protein fractions were 29kD, 24kD, 22kD, 20kD.

Electrophoretic Mobility to Monitor Protein-Surfacant Interactions

  • Hong, Soon-Taek
    • Preventive Nutrition and Food Science
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    • v.3 no.2
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    • pp.143-151
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    • 1998
  • Protein -surfactant interactions have been investigate by measuring ζ-potential of $\beta$-lactoglobulin-coated emulsion droplets and $\beta$-lactoglobulin in solution in the rpesenceof surfactant, with particular emphasis on the effect of protein heat treatment(7$0^{\circ}C$, 30min). When ionic surfactant (SDS or DATEM) is added to the protein solution, the ζ-potential of the mixture is found to increase with increasing surfactant concentration, indicating surfactant binding to the protein molecules. For heat-denatured protein,it has been observed that the ζ-potential tends to be lower than that of the native protein. The effect of surfactant on emulsions is rather complicated .With SDS, small amounts of surfactant addition induce a sharp increase in zeta potential arising from the specific interaction of surfactant with protein. With further surfacant addition, there is a gradual reductio in the ζ-potential, presumably caused by the displacement of adsorped protein (and protein-surfactant complex) from the emulsion droplet surfac by the excess of SDS molecules. At even higher surfactant concentrations, the measured zeta potential appears to increase slightly, possibly due to the formation of a surfactant measured zeta potential appears to increase slightly, possibly due to the formation of surfactant micellar structure at the oil droplet surface. This behaviour contrastswith the results of the corresponding systems containing the anionic emulsifier DATEM, in which the ζ-potential of the system is found to increase continuously with R, particularly at very low surfactant concentration. Overall, such behaviour is consisten with a combination of complexation and competitive displacement between surfactant and protein occurring at the oil-water interface. In addition, it has also been found that above the CMC, there is a time-dependent increase in the negative ζ-potential of emulsion droplets in solutions of SDS, possibly due to the solublization of oil droplets into surfactant micelles in the aqueous bulk phase.

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Metabolism of an Anionic Fluorescent Dye, 1-Anilino-8-naphthalene Sulfonate (ANS) by Rat Liver Microsomes

  • Chung, Youn-Bok;Bae, Woong-Tak;Han, Kun
    • Archives of Pharmacal Research
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    • v.21 no.6
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    • pp.677-682
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    • 1998
  • The present study was designed to examine the metabolism of 1-anilino-8-naphthalene sulfonate (ANS), an anionic compound which is transported into liver via "multispecific organ ic anion transporter", with rat hepatic microsomes. TLC analysis indicated that the fluorescent metabolites were not produced to a measurable extent, which made it possible to assess the ANS metabolism by measuring the fluorescence disappearance. The metabolism of ANS was remarkably inhibited by the presence of SKF-525A as well as by the substitution of 02 by CO gas. ANS metabolism by microsomes also required NADPH as a cofactor. These results indicated that the microsomal monooxygenase system might be mainly responsible for the ANS metabolism. The maximum velocity ($V_{max}$) and Michaelis constant ($K_m$) were calculated to be $4.3{\pm}0.2$ nmol/min/mg protein and $42.1{\pm}2.0\;{\mu}M$, respectively. Assuming that 1g of liver contains 32mg of microsomal protein, the $V_{max}$ value was extrapolated to that per g of liver ($V_{max}^I$). The intrinsic metabolic clearance ($CL_{int}$) under linear conditions calculated from this in vitro metabolic study was 3.3ml/min/g liver, being comparable with that (3.0ml/min/g liver) calculated by analyzing the in vivo plasma disappearance curve in a previous study. Furthermore, the effects of other organic anions on the metabolism of ANS were examined. Bromophenolblue (BPB) and rose bengal (RB) competitively inhibited the metabolism of ANS, while BSP inhibited it only slightly. The inhibition constant ($K_i$) of BPB ($6\;{\mu}M$) was much smaller than that of RB ($200\;{\mu}M$). In conclusion, the microsomal monooxygenase system plays a major role in the metabolism of ANS, and other unmetabolizable organic anions (BPB and RB) compete for this metabolism.

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Electrophoretic Studies on Haemolymph Protein, Digestive Fluid Protein and Digestive Amylase Activity in the Wild silkworm, Theophila mandarina. (상잠의 형액과 소화액에 있어서 단백질과 Amylase 활성에 관한 전기영동적 연구)

  • 문재유;박광의
    • Journal of Sericultural and Entomological Science
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    • v.27 no.1
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    • pp.37-41
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    • 1985
  • The haemolymph protein, digestive fluid proteins and digestive fluid amylase activity of wild silkworm, Theophila mandarina those of the were studied by polyacrylamide gel electrophoresis. In addition, they was also compared with silkworm. 1. 6 main protein bands in female and 7 main protein bands in male were detected in the larval haemolymph of T. mandarina where as 8 and 7 main protein bands in female and male of B. mori were observed. Some differences in the haemolymph protein ands of T. mandarina and B. mori were observed. 2. 15 protein bands and 12 protein bands were found in the larval digestive fluid of T. mandarina and B. mori respectively. Some differences in the mobility of digestive fluid proteins of T. mandarina and B. mori were noticed. 3. Larval digestive fluid amylases were anionic and moved near the tracking dye in both T. mandarina and B. mori. Mobility of the digestive fluid amylases relative to bromophenol blue were 0.019 and 0.020 in T. mandarina and B. mori respectively.

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Characterization of the Hepatic Uptake of l-Anilino-8-naphthalene sulfonate(ANS) by Isolated Rat Hepatocytes-Is Serum Protein Essential for Hepatic Uptake of ANS in the Liver?- (유리간세포를 사용한 ANS의 간내 이행에 관한 연구-ANS의 간내 이행과정에 단백질 매개 기구가 존재하는가?-)

  • Chung, Youn-Bok;Yuk, Dong-Yeon;Han, Kun
    • Journal of Pharmaceutical Investigation
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    • v.21 no.1
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    • pp.43-50
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    • 1991
  • The hepatic uptake of an anionic fluorescence probe, l-anilino-8-naphthalene sulfonate (ANS) was characterized using isolated rat hepatocytes. The initial uptake rate of ANS by isolated hepatocytes was determined. The uptake process of ANS was fitted well to the Michaelis-Menten equation with a saturable component. The $V_{max}$ and $K_m$ values were $2.9{\pm}0.1\;nmol/min/mg$ protein and $29.1{\pm}3.2\;{\mu}M$, respectively. The uptake clearance $(CL_{up})$ based on the ratio of $V_{max}$ to $K_m$ was 11.7 ml/min/g liver, revealing the good coincidence with that assessed from the analysis of the plasma disappearance curve in previous report. Furthermore, the effect of serum protein on the hepatic uptake of ANS into isolated hepatocytes was investigated. The permeability clearances $(PS_{inf})$ of ANS uptake were much higher than those predicted based on the unbound fractions in the presence of serum. These suggested that the hepatic uptake of extensively serum protein-bound ANS is mediated not only by the unbound form of ligand but also by the serum protein-mediated uptake mechanism.

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In situ Recovery of hGM-CSF in Transgenic Rice Cell Suspension Cultures (형질전환 벼 현탁세포 배양에서 hGM-CSF의 in situ Recovery 연구)

  • Myoung, Hyun-Jong;Choi, Hong-Yeol;Nam, Hyung-Jin;Kim, Dong-Il
    • KSBB Journal
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    • v.30 no.3
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    • pp.103-108
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    • 2015
  • Production of foreign proteins by transgenic plant cell cultures has several advantages such as post-translational modification, low risk of product contamination and low-cost production and purification. However, target proteins are degraded by extracellular proteases existing in the media. A solution to this problem is the use of perfusion culture and ion exchange chromatography for the application of integrated bioprocess using in situ recovery. With this method, production of human granulocyte-macrophage colony-stimulating factor (hGM-CSF) was investigated in this study. First, optimization of cell concentration during the induction phase for the production of hGM-CSF was examined. As cell concentration increased, the level of hGM-CSF was decreased due to the presence of extracellular proteases. Induction using sugarfree media produced 33% more hGM-CSF. The effects of pH on the binding of hGM-CSF to cationic and anionic exchange resins were also investigated. In terms of stability, optimal pH was found to be 5~7. In the case of using buffer exchange when CM-Sepharose was used as a cationic exchange resin, optimal pH for binding was 4.8 and adsorption yield was 77%. When DEAE-Sepharose was used as an anionic exchange resin, it was 5.5 (74%). Without buffer exchange, optimal pH was 4.6 (84%). From these results, an integrated bioprocess using in situ recovery with simultaneous production and separation of foreign protein in transgenic plant cell suspension cultures was found to be feasible.