• 제목/요약/키워드: binding treatment

검색결과 1,247건 처리시간 0.031초

Separation of Calcium-binding Protein Derived from Enzymatic Hydrolysates of Cheese Whey Protein

  • Kim, S.B.;Shin, H.S.;Lim, J.W.
    • Asian-Australasian Journal of Animal Sciences
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    • 제17권5호
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    • pp.712-718
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    • 2004
  • This study was carried out to separate the calcium-binding protein derived from enzymatic hydrolysates of cheese whey protein. CWPs (cheese whey protein) heated for 10 min at $100^{\circ}C$ were hydrolyzed by trypsin, papain W-40, protease S, neutrase 1.5 and pepsin, and then properties of hydrolysates, separation of calcium-binding protein and analysis of calcium-binding ability were investigated. The DH (degree of hydrolysis) and NPN (non protein nitrogen) of heated-CWP hydrolysates by commercial enzymes were higher in trypsin than those of other commercial enzymes. In the result of SDS-PAGE (sodium dodecyl sulphate polyacrylamide gel electrophoresis), $\beta$-LG and $\alpha$-LA in trypsin hydrolysates were almost eliminated and the molecular weight of peptides derived from trypsin hydrolysates were smaller than 7 kDa. In the RP-HPLC (reverse phase HPLC) analysis, $\alpha$-LA was mostly eliminated, but $\beta$-LG was not affected by heat treatment and the RP-HPLC patterns of trypsin hydrolysates were similar to those of SDS-PAGE. In ion exchange chromatography, trypsin hydrolysates were shown to peak from 0.25 M NaCl and 0.5 M NaCl, and calcium-binding ability is associated with the large peak, which was eluted at a 0.25 M NaCl gradient concentration. Based on the results of this experiment, heated-CWP hydrolysates by trypsin were shown to have calcium-binding ability.

Investigation of the Binding Site of CCR2 using 4-Azetidinyl-1-aryl-cyclohexane Derivatives: A Membrane Modeling and Molecular Dynamics Study

  • Kothandan, Gugan;Gadhe, Changdev G.;Cho, Seung Joo
    • Bulletin of the Korean Chemical Society
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    • 제34권11호
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    • pp.3429-3443
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    • 2013
  • Chemokine receptor (CCR2) is a G protein-coupled receptor that contains seven transmembrane helices. Recent pharmaceutical research has focused on the antagonism of CCR2 and candidate drugs are currently undergoing clinical studies for the treatment of diseases like arthritis, multiple sclerosis, and type 2 diabetes. In this study, we analyzed the time dependent behavior of CCR2 docked with a potent 4-azetidinyl-1-aryl-cyclohexane (4AAC) derivative using molecular dynamics simulations (MDS) for 20 nanoseconds (ns). Homology modeling of CCR2 was performed and the 4AAC derivative was docked into this binding site. The docked model of selected conformations was then utilized to study the dynamic behavior of the 4AAC enzyme complexes inside lipid membrane. MDS of CCR2-16b of 4AAC complexes allowed us to refine the system since binding of an inhibitor to a receptor is a dynamic process and identify stable structures and better binding modes. Structure activity relationships (SAR) for 4AAC derivatives were investigated and reasons for the activities were determined. Probable binding pose for some CCR2 antagonists were determined from the perspectives of binding site. Initial modeling showed that Tyr49, Trp98, Ser101, Glu291, and additional residues are crucial for 4AAC binding, but MDS analysis showed that Ser101 may not be vital. 4AAC moved away from Ser101 and the hydrogen bonding between 4AAC and Ser101 vanished. The results of this study provide useful information regarding the structure-based drug design of CCR2 antagonists and additionally suggest key residues for further study by mutagenesis.

Perilla Leaf Extract Inhibits 3T3-L1 Preadipocytes Differentiation

  • Kim, Mi-Ja;Kim, Hye-Kyung
    • Food Science and Biotechnology
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    • 제18권4호
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    • pp.928-931
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    • 2009
  • Effects of perilla leaf extracts (PLE) on adipocytes differentiation of 3T3-L1 cells were examined. Ethanol extract of PLE treatment significantly decreased lipid accumulation, a marker of adipogenesis, in a dose-dependent manner. Moreover, gene expression levels of peroxisome proliferator-activated receptor ${\gamma}$ ($PPAR{\gamma}$), the key adipogenic transcription factor, were markedly decreased by PLE. PLE also suppressed adipocyte fatty acid binding protein (aP2) and glycerol-3-phosphate dehydrogenase (GPDH), which are adipogenic marker proteins. These results suggest that PLE treatment suppressed differentiation of 3T3-L1 adipocytes, in part by down-regulating expression of adipogenic transcription factor and other specific target genes.

닭털 단백질로부터 가수분해물 제조 및 철분 결합 펩타이드의 분리 (Preparation of chicken feather protein hydrolysates and isolation of iron-binding peptides)

  • 김남호;최동원;송경빈
    • 한국식품저장유통학회지
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    • 제20권3호
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    • pp.435-439
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    • 2013
  • 닭털 단백질을 단백질 가수분해 효소인 Flavouzyme을 이용하여 8시간 동안 가수분해하여 가수분해물을 제조하였다. 닭털 단백질 가수분해물로부터 저분자량 펩타이드를 얻고자 한외여과를 하였고, 철분 결합 펩타이드를 분리하기 위해 Q-Sepharose와 Sephadex G-15 컬럼을 사용하여 분리하였다. 그 결과, 철분 결합력이 높은 펩타이드 분획, F12를 분리하였고, 향후 철분 보충제로써 활용이 가능하다고 판단된다.

콩과 토란에서 추출한 FITC-Lectin의 마우스 소장조직에 대한 현미경 관찰 (Light and Electron Microscopical Observation of the Binding of Lectin to Mouse Intestine)

  • 서영주
    • 한국식품영양과학회지
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    • 제22권4호
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    • pp.494-499
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    • 1993
  • 렉틴투여 마우스의 소장을 고정절편으로 해서, HE 염색한 후 광학현미경관찰 및 조직을 반전고정해서 주사형전자현미경관찰을 하여, 소장점막의 미융모막의 변화를 대조군과 비교했다. 그 결과 소장융모의 팽윤, 단평화, 소장벽의 박약화, 상피세포의 밀도화 및 흐트러짐 등이 관찰되었다. 즉 렉틴이 정상적인 생체기능을 방해한다는 의미에서의 독활성이 있다는 것은, 소장조직에의 영양소흡수부전이 하나의 요인이 됨을 알 수 있다.

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Comparative lectin binding patterns of Cochlodinium polykrikoides Margalef

  • Rhodes, Lesley L.;Cho, Yong-Chul;Cho, Eun-Seob
    • Journal of the korean society of oceanography
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    • 제35권3호
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    • pp.153-157
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    • 2000
  • Four different FITC-conjugated lectins were used to visually evaluate lectin binding activity by optical staining quality using confocal laser scanning microscopy (CLSM) of Cochzodinium polykrikoides in nature (wild type) and culture (cultured type). Cells from the field and cultures treated with ConA fluoresced only at the outer cell wall, and the abundance and distribution of the fluorescent signal were similar. Treatment with PWM and HPA did not elicit fluorescence at the cell surface, but the wild type exposed to HPA showed greater binding than did the cultured cells, possibly due to greater concentrations of glucosamine. The wild type cells treated with LBL lectin showed a strong green fluorescence on the cell surface, whereas cultured cells did not. Signal intensity and abundance were greater than for any other lectins tested in this study. These results suggest that wild type and cultured type are significantly different based on surface sugar production. In particular, the wild type cells apear richer in galactosamine-like moieties. Neither glucose nor mannose-like moieties were present in either wild types or cultured cells.

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Maltose Binding Protein 융합단백질에 의한 인간유래의 앤지오스타틴과 앤도스타틴의 대장균에서 수용성 단백질발현 (Soluble Expression of Human Angiostatin and Endostatin by Maltose Binding Protein (MBP) Fusion in E. coli)

  • 박선열;최신건
    • 산업기술연구
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    • 제28권B호
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    • pp.59-63
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    • 2008
  • Rapid production of therapeutic proteins such as angiostatin and endostatin angiogenic inhibititors has been highly demanded for cancer treatment. In this regard, recombinant human angiostatin and endostatin were successfully expressed as soluble forms by maltose binding protein (MBP)-mediated fusion expression in Escherichia coli. PCR amplified, angiostatin and endostatin genes from human placenta cDNA library were inserted into an expression vector pMAL-c2e to construct prokaryotic expression vectors, pMAL-c2e/AS and pMAL-c2e/ES, respectively. Recombinant angiostatin and endostatin were efficiently expressed in E. coli origami (DE3) after IPTG induction and protein expression were confirmed by SDS-PAGE analyses. The expressed recombinant proteins were purified near homogenity using an amylose affinty column chromatography. In contrast that previous E. coli expressions were all insoluble, our results first time demonstrated that MBP fused human angiostatin and endostatin were soluble in E. coli.

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Proteome Analysis of Vernalization-Treated Arabidopsis thaliana by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry

  • Cho, Mi-Ran;Lee, Kyung-Hyeon;Hyun, You-Bong;Lee, Il-Ha;Kim, Hie-Joon
    • Bulletin of the Korean Chemical Society
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    • 제28권3호
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    • pp.427-431
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    • 2007
  • In order to gain insight into the molecular changes at the protein level in plants exposed to low temperature for a long period of time (vernalization), proteome analyses of vernalization-treated Arabidopsis thaliana have been carried out by two-dimensional gel electrophoresis followed by matrix-assisted laser desorption/ ionization time-of-flight mass spectrometry. Fourteen proteins including ATP binding/GTP binding/translation elongation factor and glycine-rich RNA-binding protein 7 (GRP7) showed differential expression in vernalization-treated Arabidopsis thaliana. GRP7 showed the most dramatic increase in expression suggesting its involvement in response to vernalization treatment.

옥수수(Zea mays L.) 자엽초 절편에서 Naphtylphthalamic Acid에 대한 오옥신 이동계의 감지적응 (Sensory Adaptation in Polar Auxin Transport System to Naphtylphthalamic Acid in Corn Coleoptile Segments)

  • 윤인선
    • Journal of Plant Biology
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    • 제34권4호
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    • pp.317-323
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    • 1991
  • Partial recovery in auxin transport capacity from inhibition by N-naphthylphthalamic acid (NPA) was observed when corn coleoptile segments were subjected to a prolonged NPA treatment. Kinetic data indicated that the recovery time is a function of the concentration of NPA applied. Desensitization to NPA was also seen in tissue slices where NPA increased net uptake of auxin, indicating that the apparant adaptation in the auxin transport system did not results possibly from auxin accumulated during transport inhibition. Studies on in vitro binding of NPA to membrane vesicles isolated from the coleoptile indicated that preincubation of the tissue with NPA resulted in the reduced binding activity. Scatchard analysis of the data indicated that this was due to decreases in the number of NPA binding sites. The possibility of causal relationship of modified NPA receptors to the sensory adaptation in auxin transport observed in coleoptile segments will be discussed.

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RNA-Binding Proteins in Amyotrophic Lateral Sclerosis

  • Zhao, Melody;Kim, Jihye Rachel;van Bruggen, Rebekah;Park, Jeehye
    • Molecules and Cells
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    • 제41권9호
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    • pp.818-829
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    • 2018
  • Significant research efforts are ongoing to elucidate the complex molecular mechanisms underlying amyotrophic lateral sclerosis (ALS), which may in turn pinpoint potential therapeutic targets for treatment. The ALS research field has evolved with recent discoveries of numerous genetic mutations in ALS patients, many of which are in genes encoding RNA binding proteins (RBPs), including TDP-43, FUS, ATXN2, TAF15, EWSR1, hnRNPA1, hnRNPA2/B1, MATR3 and TIA1. Accumulating evidence from studies on these ALS-linked RBPs suggests that dysregulation of RNA metabolism, cytoplasmic mislocalization of RBPs, dysfunction in stress granule dynamics of RBPs and increased propensity of mutant RBPs to aggregate may lead to ALS pathogenesis. Here, we review current knowledge of the biological function of these RBPs and the contributions of ALS-linked mutations to disease pathogenesis.