• Title/Summary/Keyword: Binding property

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Identification of Ligand for Salivary Lipocalin Secreted from the Uterine Endometrium during Early Pregnancy in Pigs

  • Seo, Hee-Won;Kim, Min-Goo;Ka, Hak-Hyun
    • Journal of Embryo Transfer
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    • v.24 no.4
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    • pp.259-263
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    • 2009
  • Salivary lipocalin (SAL1) is a member of the lipocalin protein family that has a property to associate with many lipophilic molecules and was identified as pheromone-binding protein in pigs. Our previous study has shown that SAL1 is expressed in the uterine endometrium in a cell type- and implantation stage-specific manner and secreted into the uterine lumen in pigs. However, function of SAL1 in the uterus during pregnancy in pigs is still not known. To understand physiological function of SAL1 in the uterine endometrium during pregnancy in pigs, it needs to elucidate the ligand(s) for SAL1. Thus, to identify the ligand for SAL1 in the porcine uterus, we collected uterine luminal fluid from pigs on day 12 of pregnancy by flushing with PBS. Proteins from the uterine luminal fluid were separated by ion exchange chromatography and gel filtration. Fractions containing SAL1 protein were pooled and concentrated. Immunoblot analysis confirmed successful purification of SAL1. Then, we extracted lipids from the purified SAL1 protein and analyzed the lipids by liquid chromatography-mass spectrometry, and predicted to be steroid hormones and prostaglandins as SAL1 ligands. Results in this study showed that SAL1 protein in the uterine secretions has a small lipophilic molecule as a natural ligand. Further characterization of ligand extracted from purified SAL1 will be useful for understanding physiological function of SAL1 during pregnancy and its application to increase the pregnancy rate in pigs.

Comparative Dynamics of $tRNA^{val}$ and pBluescript II SK(+) Phagemid Studied with Ethidium Bromide and a Long-lifetime Metal-ligand Complex

  • Kang, Jung-Sook;Yoon, Ji-Hye
    • Journal of Photoscience
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    • v.11 no.3
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    • pp.133-139
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    • 2004
  • The metal-ligand complex, $[Ru(phen)_2(dppz)]^{2+}$ (phen=1,10-phenanthroline, dppz=dipyrido[3,2-a:2',3'-c]phenazine) (RuPD), was used as a spectroscopic probe for studying nucleic acid dynamics. The RuPD complex displays a long lifetime and a molecular light switch property upon DNA binding due to shielding of its dppz ligand from water. To show the usefulness of this luminophore (RuPD) for probing nucleic acid dynamics, we compared its intensity and anisotropy decays when intercalated into the $tRNA^{val}$ and pBluescript (pBS) II SK(+) phagemid through a comparison with ethidium bromide (EB), a conventional nucleic acid probe. We used frequency-domain fluorometry with a blue light-emitting diode (LED) as the modulated light source. The mean lifetime for the $tRNA^{val}$ (<${\tau}$> = 166.5 ns) was much shorter than that for the pBS II SK(+) phagemid (<${\tau}$> = 481.3 ns), suggesting a much more efficient shielding from water by the phagemid. Because of their size difference, the anisotropy decay data showed a much shorter rotational correlation times for the $tRNA^{val}$ (99.9 and 23.6 ns) than for the pBS II SK(+) phagemid (968.7 and 39.5 ns). These results indicate that RuPD can be useful for studying nucleic acid dynamics.

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Gelatinization Properties and Molecular Structure of Waxy Rice Starches Isolated from Korean Japonica and Indica Cultivars (국내산 자포니카와 인디카 품종 찹쌀전분의 호화특성과 분자구조)

  • Oh, Song Min;No, Jun Hee;Shin, Malshick
    • Korean journal of food and cookery science
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    • v.30 no.6
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    • pp.716-725
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    • 2014
  • Gelatinization properties and the molecular structure of Korean waxy rice starchesisolated from two japonica types, Sinseonchal, and Dongjinchal, as well as an indica type, Hangangchal 1 were investigated. Sinseonchal is preferred cultivar for making Korean traditional rice cakes and cookies. Sinseonchal starch was the highest in crude protein, amylopectin, damaged starch contents, and water binding capacity among the cultivars tested. The initial pasting temperature ($72.75^{\circ}C$), peak (360.54 RVU), breakdown (162.21 RVU) and setback (30.83 RVU) viscosities of Sinseonchal had the highest values (p<0.05). Onset and peak temperatures by differential scanning calorimeter were also the highest in Sinseonchal. The molecular weight of Sinseonchal amylopectin was 5.46 107higher than those of the other cultivars, but its peak height and area were the lowest among them. The amylopectin peak by HPSEC showed a shoulder in the lower molecular weight portion and its relative area decreased in the following order; Sinseonchal > Dongjinchal > Hangangchal 1. On the branch chain length distribution of amylopectin, the proportion of DP13-24 and DP25-36 showed reverse trends, with higher japonica type amylopectin in DP13-24.

Visualization of Geometric Features in the Contact Region of Proteins (단백질 접촉 영역의 기하학적 특성 가시화)

  • Kim, Ku-Jin
    • KIPS Transactions on Software and Data Engineering
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    • v.8 no.10
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    • pp.421-426
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    • 2019
  • In this paper, we propose a method to visualize the geometric features of the contact region between proteins in a protein complex. When proteins or ligands are represented as curved surfaces with irregularities, the property that the two surfaces contact each other without intersections is called shape compatibility. Protein-Protein or Protein-Ligand docking researches have shown that shape complementarity, chemical properties, and entropy play an important role in finding contact regions. Usually, after finding a region with high shape complementarity, we can predict the contact region by using residual polarity and hydrophobicity of amino acids belonging to this region. In the research for predicting the contact region, it is necessary to investigate the geometrical features of the contact region in known protein complexes. For this purpose, it is essential to visualize the geometric features of the molecular surface. In this paper, we propose a method to find the contact region, and visualize the geometric features of it as normal vectors and mean curvatures of the protein complex.

Isolation and Characterization of an Eosinophilic GH 16 β-Agarase (AgaDL6) from an Agar-Degrading Marine Bacterium Flammeovirga sp. HQM9

  • Liu, Yan;Tian, Xiaoxu;Peng, Chao;Du, Zongjun
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.235-243
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    • 2019
  • A special eosinophilic agarase exo-type ${\beta}$-agarase gene, AgaDL6, was cloned from a marine agar-degrading bacterium, Flammeovirga sp. HQM9. The gene comprised 1,383-bp nucleotides encoding a putative agarase AgaDL6 of 461 amino acids with a calculated molecular mass of 52.8 kDa. Sequence analysis revealed a ${\beta}$-agarase domain that belongs to the glycoside hydrolase family (GH) 16 and a carbohydrate-binding module (CBM_4_9) unique to agarases. AgaDL6 was heterologously expressed in Escherichia coli BL21 (DE3). Enzyme activity analysis of the purified protein showed that the optimal temperature and pH of AgaDL6 were $50^{\circ}C$ and 3.0, respectively. AgaDL6 showed thermal stability by retaining more than 98% of activity after incubation for 2 h at $50^{\circ}C$, a feature quite different from other agarases. AgaDL6 also exhibited outstanding acid stability, retaining 100% of activity after incubation for 24 h at pH 2.0 to 5.0, a property distinct from other agarases. This is the first agarase characterized to have such high acid stability. In addition, we observed no obvious stimulation or inhibition of AgaDL6 in the presence of various metal ions and denaturants. AgaDL6 is an exo-type ${\beta}$-1,4 agarase that cleaved agarose into neoagarotetraose and neoagarohexaose as the final products. These characteristics make AgaDL6 a potentially valuable enzyme in the cosmetic, food, and pharmaceutical industries.

Flavonoid Silibinin Increases Hair-Inductive Property Via Akt and Wnt/β-Catenin Signaling Activation in 3-Dimensional-Spheroid Cultured Human Dermal Papilla Cells

  • Cheon, Hye In;Bae, Seunghee;Ahn, Kyu Joong
    • Journal of Microbiology and Biotechnology
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    • v.29 no.2
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    • pp.321-329
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    • 2019
  • Hair loss, also known as alopecia, is a common dermatological condition of psychosocial significance; development of therapeutic candidates for the treatment of this condition is, hence, important. Silibinin, a secondary metabolite from Silybum marianum, is an effective antioxidant that also prevents various cutaneous problems. In this study, we have investigated the effect of silibinin on hair induction using three-dimensional (3D) cultured, human dermal papilla (DP) spheroids. Silibinin was found to significantly increase viability through AKT serine/threonine kinase (AKT) activation in 3D DP spheroids. This was correlated with an increase in the diameter of the 3D DP spheroids. The activation of the wingless and INT-1 (Wnt)/${\beta}$-catenin signaling pathway, which is associated with hair growth induction in the DP, was evaluated using the T cell-specific transcription factor and lymphoid enhancer-binding factor (TCF/LEF) transcription factor reporter assay; results indicated significantly increased luciferase activity. In addition, we were able to demonstrate increased expression of the target genes, WNT5a and LEF1, using quantitative real-time PCR assay. Lastly, significantly elevated expression of signature genes associated with hair induction was demonstrated in the 3D DP spheroids treated with silibinin. These results suggest that silibinin promotes proliferation and hair induction through the AKT and Wnt/${\beta}$-catenin signaling pathways in 3D DP spheroids. Silibinin can be a potential candidate to promote hair proliferation.

Surface Modification of Nanofiltration Membrane with Silane Coupling Agents for Separation of Dye (실란 표면 개질된 나노복합막의 염료 분리 특성)

  • Park, Hee Min;Lim, Jee Eun;Kim, Seong Ae;Lee, Yong Taek
    • Membrane Journal
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    • v.28 no.6
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    • pp.414-423
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    • 2018
  • In this study, the commercial nanofiltration membranes were modified with octyltrimethoxysilane(OcTMS) and (3-aminopropyl)trimethoxysilane (APTMS) to improve fouling resistance and to separate dye. The chemical structure and binding energy of elements of silane-deposited surface were analyzed using XPS analysis. And the morphology and hydrophilicity property of silane-modified NF membrane were analyzed using FE-SEM, EDX, AFM, and contact angle. The surface charge of silane-modified NF membrane was characterized by zeta potentiometer analyzer. As a result, silane-modified NF membrane improved fouling resistance about 2 times as compared with that of the commercial membrane. And the silane-modified NF membranes effectively were removed cation dye over 98%.

Inhibitory effects of Porphyra dentata extract on 3T3-L1 adipocyte differentiation

  • Choi, Su-Young;Lee, Su Yeon;Jang, Da hye;Lee, Suk Jun;Cho, Jeong-Yong;Kim, Sung-Hak
    • Journal of Animal Science and Technology
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    • v.62 no.6
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    • pp.854-863
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    • 2020
  • This study was aimed to investigate the inhibitory effects of Porphyra dentata (P. dentata) extract on the adipogenesis of 3T3-L1 cells and evaluate its anti-obesity effect. The proliferation of 3T3-L1 cells and differentiation of adipocytes under treatment of P. dentata extract was examined by measuring the cell viability using alamarBlue assay and lipid droplets by Oil Red O staining. Results showed that P. dentata extract has no cytotoxicity effect and lipid droplets formation decreased in a concentration-dependent manner in 3T3-L1 cells. It has been confirmed that transcription factors affecting lipid accumulation and anti-adipogenic effects during cell differentiation are linked to P. dentata extract. We observed that P. dentata shows lowering the mRNA expression of peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα) that adipogenesis-associated key transcription factors and inhibiting adipogenesis in the early stages of differentiation. Treating the cells with P. dentata did not only suppressed PPARγ2 and C/EBPα but also significantly decreased the mRNA expression of adiponectin, Leptin, fatty acid synthase, adipocyte protein 2, and Acetyl-coA carboxylase 1. Overall, the P. dentata extract demonstrated inhibitory property in adipogenesis, which has a potential effect in anti-obesity in 3T3-L1 cells.

Variation of Antimicrobial Peptide in the Extract of the Hard-shelled Mussel Mytilus coruscus Depending on Boiling (가열 유무에 따른 참담치(Mytilus coruscus) 추출물 내의 항균 펩타이드 변화)

  • Lee, Ji-Eun;Seo, Jung-Kil
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.55 no.6
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    • pp.875-885
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    • 2022
  • This study was performed to confirm the optimal extraction method for antimicrobial peptides from the Hard-shelled mussel. Extractions were performed with two processes including 1% HAc/boiling and 1% HAc/non-boiling methods and used extracts for the comparison of the antimicrobial activity, protease stability, action mechanism, AU-PAGE (acid-urea PAGE), and HPLC chromatograms. 1% HAc/boiling extract showed potent antibacterial activities both against Gram-positive and negative bacterium but 1% HAc/non-boiling extract showed antibacterial activity only against Gram-positive bacteria. Treatment of 1% HAc/boiling extract with proteases retained almost antibacterial activity against B. subtilis, but abolished significant antibacterial activity against E. coli D31. Only 1% HAc/boiling extract showed two discrete clearing antibacterial zones including slow migrating and rapid migrating zones. Both extracts showed strong DNA-binding ability but did not show bacterial membrane permeabilizing ability. In comparison of the chromatogram obtained from C18 or cation-exchange HPLC, the eluted peaks from 1% HAc/boiling extract showed high hydrophobic property or absorbance compared to 1% HAc/non-boiling extract, respectively. The concentration of the purified antimicrobial peptide was also higher in 1% HAc/boiling extract than in 1% HAc/non-boiling extract. Our results suggest that the effective extraction condition for antimicrobial peptides from marine invertebrate is boiling process in a weak acetic acid solution (1%).

Physicochemical Properties and Storage Stability of Plant-based Alternative Meat Products Prepared with Low-Fat Soybean Powder Treated by Supercritical CO2 (초임계 이산화탄소 처리된 저지방 대두분말로 제조한 식물성 대체육의 이화학적 품질 특성 및 저장 안정성)

  • Min-Jeoung Pyo;Kyo-Yeon Lee;Chae-Yeon Han;Chae-Eun Park;Sung-Gil Choi
    • The Korean Journal of Food And Nutrition
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    • v.36 no.4
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    • pp.264-273
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    • 2023
  • Physicochemical properties and storage stability of plant-based alternative meat prepared with low-fat soybean powder (LPAM) treated by supercritical-CO2 and those of full-fat soybean powder (FPAM) were compared. Ash and crude protein contents were higher in LPAM than in FRAM. Water absorption capacity and oil absorption capacity were significantly higher in LPAM than in FPAM. Water binding capacity was higher in LPAM than in FPAM during a 20 days storage period at 5℃ and pH was significantly lower in LPAM than in FPAM after a 5~10 days storage period. Hardness, gumminess and chewiness significantly increased with the increase in the storage period, and the three were significantly higher in LPAM than in FPAM after 10 days and 20 days of storage. The acid value showed no remarkable difference according to the storage period in LPAM; however, it was significantly higher in FPAM than in LPAM after 20 days of storage. The peroxide value and TBA value were significantly increased according to the storage period, and were significantly lower iin LPAM than in FPAM during all the storage periods. Therefore, the use of low-fat soybean powder may be effective in improving oxidative stability during storage in the production of plant-based alternative meat.