• Title/Summary/Keyword: Membrane protein

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INTERACTION OF TENECIN FRAGMENTS WITH LIPOSOMES

  • Park, Myeong-Jun;Cho, Hyun-Sook;Hong, Sung-Yu;Yoon, Jeong-Hyeok;Lee, Keun-Hyeong;Moon, Hong-Mo;Cheong, Hong-Seok
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.37-37
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    • 1996
  • Tenecin fragments are antimicrobial and antifungal peptide from Tenebrio molitor with highly positive charged amino acid residues. To elucidate their membrane selectivity and molecular mechanism, various forms of tenecin fragments were synthesized, and their interaction with acidic phospholipid, Gram (+), fungal and human erythrocyte membrane were investigated by ANTS/DPX leakage, membrane binding and fusion assay. (omitted)

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Biophysical effect of lipid modification at palmitoylation site on the structure of Caveolin 3

  • Ma, Yu-Bin;Kang, Dong-Hoon;Kim, Myeongkyu;Kim, Ji-Hun
    • Journal of the Korean Magnetic Resonance Society
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    • v.23 no.3
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    • pp.67-72
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    • 2019
  • Caveolae are small plasma membrane invaginations that play many roles in signal transduction, endocytosis, mechanoprotection, lipid metabolism. The most important protein in caveolae is the integral membrane protein, caveolin, which is divided into three families such as caveolin 1, caveolin 2, and caveolin 3. Caveolin 1 and 3 are known to incorporate palmitate through linkage to three cysteine residues. Regulation of the protein palmitoylation cycle is important for the cellular processes such as intracellular localization of the target protein, membrane association, conformation, protein-protein interaction, and activity. However, the detailed aspect of individual palmitoylation has not been studied. In the present work, the role of each lipid modification at three cysteines was studied by NMR. Our results suggest that each lipid modification at the natively palmitoylation site has its own roles. For example, lipidations to C106 and C129 are play a role in structural stabilization, however, interestingly, lipid modification to C116 interrupts the structural stabilization.

Electrophoretic analysis of the major protein of erythrocyte membrane in man, bovine, horse, and dog: their relation to erythrocyte sedimentation rate (사람, 소, 말, 개의 적혈구막 단백의 전기 영동법에 의한 분석 - 적혈구 침강 속도와의 관계 -)

  • Bahk, Yeong-woo
    • Korean Journal of Veterinary Research
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    • v.41 no.1
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    • pp.21-28
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    • 2001
  • The protein of the bovine, horse and dog erythrocyte membrane were analyzed by polyacrylamide gel eletrophoresis in sodium dodecyl sulfate and their relation to the sedimentation rate of animal erythrocytes were investigated by treating the erythrocytes with proteinases such as trypsin and chymotrypsin. Protein content in erythrocyte membrane was in human, in Jindo dog, in cattle and in horse, showing similar in among. The erythrocyte sedimentation rates bovine erythrocytes from Hostein and Korean native cattle were very slow compared with the human one(1/7 as slow as the human one) as reported previously. Although the general protein profiles of the bovine erythrocyte membranes were almost similar to that of human, bovine erythrocyte membranes showed one additional protein band, called band Q in this study, which migrated electrophoretically to the mid-position between band 2 and band 3 in human erythrocyte membranes. The erythrocyte sedimentation of race horse were very fast compared with the human one are reported previously. Although the general protein profiles of the race horse erythrocyte membranes were almost similar to that of human, band 3 content was showing higher in race horse(34.7%) than in human(25.3%). The general protein profile of the Jindo dog erythrocyte membrane was almost similar to the human patterns, Jindo dog erythrocyte membranes showed one absent protein band. It was band 7. The glycoprotein profiles of the bovine erythrocyte membranes revealed by periodic acid-Schiff(PAS) stain showed a marked difference from that of human. The PAS-1(glycophorin) and PAS-2(sialoglycoprotein) present in human erythrocyte membrane were almost absent from the bovine erythrocyte membranes showed a strong PAS-positive band near the origin of the electraphorograms, which is named as PAS-B in this study. The PAS-1 and PAS-2 present in human erythrocyte membrane were almost absent from race horse erythrocyte membranes, but PAS-2 was more in only race horse from that of human. The PAS-1 and PAS-2 were absolutely absent from the Jindo dog erythrocyte membrane. These results suggest the slow sedimentation rate of bovine erythrocytes is due in part to the presence of band Q protein fraction and PAS-B glycoprotein in the bovine erythrocytes, and that the fast sedimentation rate of race horse erythrocyte is due in part to the presence of more band 3 protein fraction and PAS-E glycoproteins in the race horse erythrocytes.

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Affinity Filtration Chromatography of Proteins by Chitosan and Chitin Membranes: 2. Separation of BSA and Lysozyme (키토산 및 키틴 막에 의한 단백질의 친화 여과 크로마토그래피: 2. BSA 및 Lysozyme의 분리)

  • Youm, Kyung-Ho;Yuk, Yeong-Jae
    • Membrane Journal
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    • v.19 no.2
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    • pp.113-121
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    • 2009
  • Porous affinity chitosan and chitin membranes with good mechanical strength and high protein binding capacity were prepared by using silica particles as porogen. The maximum binding capacity of affinity chitosan membrane for BSA protein is 21.8mg/mL, and that of affinity chitin membrane for lysozyme enzyme is 26.1mg/mL. Chromatographic separations of BSA and lysozyme proteins using the porous affinity chitosan and chitin membranes were performed with change of the flow rate, loading amount and concentration of protein loading solutions. Protein eluted amount and binding yield were calculated from the filtration chromatograms consisted of loading/washing/elution sequences. Protein binding amount and yield were increased with decreasing of flow rate, increasing of loading amount and concentration of protein loading solutions. Those results suggest that the porous chitosan and chitin membranes prepared by using silica particles as porogen are suitable in affinity filtration chromatography for large scale separation of proteins.

Characterization of membrane fouling and CEB (Chemical enhanced backwashing) efficiency with temperature in SMBR Process (MBR 공정에서 수온에 따른 막오염 및 CEB 세정효율 특성)

  • Park, Kitae;Park, Jeonghoon;Choi, Eunhye;Kim, Hyungsoo;Kim, Jihoon
    • Journal of Korean Society of Water and Wastewater
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    • v.31 no.5
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    • pp.389-395
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    • 2017
  • In this paper, we investigate the characteristics of membrane fouling caused by water temperature in the Membrane bioreactor(MBR) process and try to derive the membrane fouling control by chemical enhanced backwashing(CEB). The extracellular polymeric substances(EPS) concentration was analyzed according to the water temperature in the MBR, and the membrane fouling characteristics were investigated according to the conditions, with sludge & without sludge, through a lab-scale reactor. As shown in the existing literature the fouling resistance rate was increased within sludge with the water temperature was lowered. However, in the lab-scale test using the synthetic wastewater, the fouling resistance increased with the water temperature. This is because that the protein of the EPS was more easily adsorbed on the membrane surface due to the increase of entropy due to the structural rearrangement of the protein inside the protein as the water temperature increases. In order to control membrane fouling, we tried to derive the cleaning characteristics of CEB by using sodium hypochlorite(NaOCl). We selected the condition with the chemicals and the retention time, and the higher the water temperature and the chemical concentration are the higher the efficiencies. It is considered that the increasing temperature accelerated the chemical reaction such as protein peptide binding and hydrolysis, so that the attached proteinaceous structure was dissolved and the frequency of the reaction collision with the protein with the chemical agent becomes higher. These results suggest that the MBRs operation focus on the fouling control of cake layer on membrane surface in low temperatures. On the other hand, the higher the water temperature is the more the operation strategies of fouling control by soluble EPS adsorption are needed.

Enzymatic Properties of a Fusion Protein between Human Cytochrome P450 1A1 and Rat NADPH-P450 Reductase Expressed in Escherichia Coli (대장균에서 발현된 인간 Cytochrome P450 1A1과 Rat NADPH-P450 Reductase와의 Fusion Protein의 효소 특성 연구)

  • 천영진;정태천;이현걸;한상섭;노정구
    • Toxicological Research
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    • v.12 no.2
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    • pp.155-161
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    • 1996
  • The enzymatic properties for NADPH-P450 reductase domain of a fusion protein between human cytochrome P450 1A1 and rat NADPH-P450 reductase expressed in Escherichia coli were investigated. The fusion plasmid pCW/1A1OR-expressed E. coli membrane showed high NADPH-cytochrome c reductase activity ($830.1\pm 85.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$), while pCW control vector and P 450 1A1 expression vector pCW/1A1 showed relatively quite low activity ($4.35\pm 0.49, 3.27\pm 0.50 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively). The kinetic curves for NADPH-cytochrome c reductase followed typical Michaelis-Menten kinetics. The $K_{max}$ and $V_{max}$ for NADPH-dependent reductase activity were $8.24\pm 2.61\mu $and $817.9\pm 60.8 nmol\cdot min^{-1}\cdot mg protein^{-1}$, respectively, whereas those for cytochrome c-dependent reductase activity were $19.97\pm 2.86\mu M$ and $1303.5\pm 67.1 nmol\cdot min^{-1}\cdot mg protein^{-1}$. The reductase activities were also compared with those of rat, porcine and human liver microsomes. The activity of pCW/ 1A1OR-expressed E. coli membrane was 15.2-fold higher than that of rat liver microsome. Treatment with benzo(a)pyrene, 7-ethoxyresorufin and $\alpha$-naphthofiavone which are known as specific substrates or inhibitor for human P450 1A1 increased NADPH-cytochrome c reductase activity of fusion protein in E. coli membrane dose-dependently. These results demonstrate that the membrane topology of fused enzyme may be important for activity of its NADPH-P450 reductase domain.

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Effect of Sexual Pheromone on Phosphoryation of Membrane Protein in Heterobasidiomycetous Yeast Rhodosporidium toruloides (이담자효모 Rhodosporidium toruloides의 막단백질 인산화와 성 Pheromone, Rhodotorucine A의 작용)

  • 정영기
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.6
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    • pp.1246-1251
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    • 1997
  • When the membrane protein fraction of mating type a cells of heterobasidiomycetous yeast R. toruloides was phosphorylated in vitro, two phosphorylated proteins of 72Kd and 57Kd were detected on SDS-polyacryamide gel. The phosphorylation reaction was inhibited by rhodotorucine A(Rh. A) which is a sexual pheromone secreted by mating type A cells. The inhibition of phosphorylation by Rh. A was dependent on $Ca^{2+}$, and independent on $Mg^{2+}$ or calmodulin. When adding trigger peptidase(TPase) inhibitor, antipain, no inhibition of phosphory was observed. Also, by adding the trysin-digested product of Rh. A, the phosphorylation was inhibited as the action of Rh. A. From these results, it is expected that the inhibition of membrane protein phosphorylation should be caused by the digested product of Rh. A with TPase.

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A Possible Significance in Vertebrate Phototransduction of Multi-Protein Signaling Complexes on Raft-Like Membranes

  • Hayashi, Fumio;Liu, Han;Seno, Keiji
    • Journal of Photoscience
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    • v.9 no.2
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    • pp.47-50
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    • 2002
  • Raft is a distinctive membrane domain enriched in a certain class of lipids, cholesterol, and proteins observed on the plasma membrane. Growing evidence has revealed that such membrane domains play key roles in signal transduction, fertilization, development, transmitter release, and so on. Recently, we have isolated raft-like detergent-resistant membrane (DRM) fraction from bovine photoreceptor rod outer segments. Transducin and its effecter, cGMP-phosphodiesterase, elicited stimulus-dependent translocation between detergent-soluble membrane and DRM. This suggested potential importance of such distinct membrane domains in vertebrate phototransduction. Here, we will discuss physiological meaning of the translocation of major components of cGMP cascade to raft-like membrane in phototransduction. We would like to propose a hypothesis that raft-like membrane domains on the disk membrane are the place where cGMP cascade system could be quenched.

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The Effects of Dietary Proteins on Hepatic Histological Changes and Membrane Stability in Chemically Induced Rat Hepatocarcinogenesis (화학적으로 유도한 랫드 간세포 암화과정에서 단백질 식이가 간의 조직학적 변화와 생체막 안정도에 미치는 영향)

  • 박경애;김현덕;최혜미
    • Journal of Nutrition and Health
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    • v.34 no.8
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    • pp.833-842
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    • 2001
  • The purpose of this study is to determine the effect of dietary proteins and fats on the hepatic histological changes, membrane stability, and drug-metabolizing enzyme activities during chemically induced rat hepatocarcinogenesis. Weanling Sprague-Dawley rats were fed the diet containing 20% casein or soy protein isolate and 15% perilla or corn oil for 10 weeks. Hepatocarcinogensis was initiated with diethylnitrosamine(DEN), and the rats were fed diets containing 0.02% 2-acetylaminofluorene(AAF) followed by 0.05% phenobarbital (PB). The scores of histological changes were decreased in treated rats fed soy protein diet compared to those find casein diet. Liver weights were significantly increased by AAF and PB treatment in rats fed casein diets in both oil groups. Glucose 6-phosphatase(G6Pase) activities, an index of membrane stability, were significantly reduced by AAF and PB treatment in rats find casein diets, and were lower in casein diet compared to soy protein diet groups. Especially, the activities were the highest in the rats fed soy protein-perilla oil diet. Lipid peroxide values also were increased by AAF and PB treatment in rats fed casein diet. Aniline hydroxylase activities were not influenced by protein and fat sources. Glutathione-dependent enzyme activities were increased by AAF and PB treatment. Linoleic and arachidonic acid content were increased in rats fed corn oil diet, and linolenic and eicosapentaenoic acid contents were increased in rats fed perilla oil diet. Our results suggest that soy protein isolate inhibit the abnormal histological changes in liver, possibly by maintaining the membrane stability during chemically induced rat hepatocarcinogenesis. Soy protein may be protective against the hepatocarcinogenesis induced by chemical carcinogen.

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Backbone 1H, 15N, and 13C Resonance Assignment of HP1242 from Helicobacter pylori

  • Kang, Su-Jin;Park, Sung-Jean;Jung, Seo-Jeong;Lee, Bong-Jin
    • BMB Reports
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    • v.38 no.5
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    • pp.591-594
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    • 2005
  • One of the small proteins from Helicobacter pylori, HP1242, was investigated by the solution nuclear magnetic resonance (NMR) spectroscopy. HP1242 is known as a 76-residue conserved hypothetical protein and its function cannot be identified based on sequence homology. Here, the results of the backbone $^1H$, $^{15}N$, and $^{13}C$ resonance assignments of the HP1242 are reported using double- and triple-resonance techniques. About 95% of all of the $^1HN$, $^{15}N$, $^{13}CO$, $^{13}C{\alpha}$, and $^{13}C{\beta}$ resonances that cover 75 non- Proline residues of the 76 residues are clarified through sequential- and specific- assignments. In addition, three helical regions were clearly identified on the basis of the resonance assignments.