• Title/Summary/Keyword: Binding Potential

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Cadmium-Substituted Concanavalin A and Its Trimeric Complexation

  • Park, Yeo Reum;Kim, Da Som;Lee, Dong-Heon;Kang, Hyun Goo;Park, Jung Hee;Lee, Seung Jae
    • Journal of Microbiology and Biotechnology
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    • v.28 no.12
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    • pp.2106-2112
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    • 2018
  • Concanavalin A (ConA) interacts with carbohydrates as a lectin, and recent reports proposed its application for detecting a diversity of viruses and pathogens. Structural studies have detailed the interaction between ConA and carbohydrates and the metal coordination environment with manganese and calcium ions (Mn-Ca-ConA). In this study, ConA was crystallized with a cadmium-containing precipitant, and the refined structure indicates that $Mn^{2+}$ was replaced by $Cd^{2+}$ (Cd-Ca-ConA). The structural comparison with ConA demonstrates that the metal-coordinated residues of Cd-Ca-ConA, that is Glu8, Asp10, Asn14, Asp19, and His24, do not have conformational shifts, but residues for sugar binding, including Arg228, Tyr100, and Leu99, reorient their side chains, slightly. Previous studies demonstrated that excess cadmium ions can coordinate with other residues, including Glu87 and Glu183, which were not coordinated with $Cd^{2+}$ in this study. The trimeric ConA in this study coordinated $Cd^{2+}$ with other residues, including Asp80 and Asp82, for complex generation. The monomer does not have specific interaction near interface regions with the other monomer, but secondary cadmium coordinated with two aspartates (Asp80 and Asp82) from monomer 1 and one aspartate (Asp16) from monomer 2. This study demonstrated that complex generation was induced via coordination with secondary $Cd^{2+}$ and showed the application potential regarding the design of complex formation for specific interactions with target saccharides.

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.

The identification of novel regions for reproduction trait in Landrace and Large White pigs using a single step genome-wide association study

  • Suwannasing, Rattikan;Duangjinda, Monchai;Boonkum, Wuttigrai;Taharnklaew, Rutjawate;Tuangsithtanon, Komson
    • Asian-Australasian Journal of Animal Sciences
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    • v.31 no.12
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    • pp.1852-1862
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    • 2018
  • Objective: The purpose of this study was to investigate a single step genome-wide association study (ssGWAS) for identifying genomic regions affecting reproductive traits in Landrace and Large White pigs. Methods: The traits included the number of pigs weaned per sow per year (PWSY), the number of litters per sow per year (LSY), pigs weaned per litters (PWL), born alive per litters (BAL), non-productive day (NPD) and wean to conception interval per litters (W2CL). A total of 321 animals (140 Landrace and 181 Large White pigs) were genotyped with the Illumina Porcine SNP 60k BeadChip, containing 61,177 single nucleotide polymorphisms (SNPs), while multiple traits single-step genomic BLUP method was used to calculate variances of 5 SNP windows for 11,048 Landrace and 13,985 Large White data records. Results: The outcome of ssGWAS on the reproductive traits identified twenty-five and twenty-two SNPs associated with reproductive traits in Landrace and Large White, respectively. Three known genes were identified to be candidate genes in Landrace pigs including retinol binding protein 7, and ubiquitination factor E4B genes for PWL, BAL, W2CL, and PWSY and one gene, solute carrier organic anion transporter family member 6A1, for LSY and NPD. Meanwhile, five genes were identified to be candidate genes in Large White, two of which, aldehyde dehydrogenase 1 family member A3 and leucine rich repeat kinase 1, associated with all of six reproduction traits and three genes; retrotransposon Gag like 4, transient receptor potential cation channel subfamily C member 5, and LHFPL tetraspan subfamily member 1 for five traits except W2CL. Conclusion: The genomic regions identified in this study provided a start-up point for marker assisted selection and estimating genomic breeding values for improving reproductive traits in commercial pig populations.

2-DE and MALDI-TOF MS-based identification of bovine whey proteins in milk collected soon after parturition

  • Lee, Jae Eun;Lin, Tao;Kang, Jung Won;Shin, Hyun Young;Lee, Joo Bin;Jin, Dong Il
    • Korean Journal of Agricultural Science
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    • v.45 no.4
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    • pp.635-643
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    • 2018
  • Bovine milk is widely consumed by humans and is a primary ingredient of dairy foods. Proteomic approaches have the potential to elucidate complex milk proteins and have been used to study milk of various species. Here, we performed a proteomic analysis using 2-dimensional electrophoresis (2-DE) and matrix assisted laser desorption ionization-time of flight mass spectrometer (MALDI-TOF MS) to identify whey proteins in bovine milk obtained soon after parturition (bovine early milk). The major casein proteins were removed, and the whey proteins were analyzed with 2-dimensional polyacrylamide gel electrophoresis (2-D PAGE). The whey proteins (2 mg) were separated by pI and molecular weight across pH ranges of 3.0 - 10.0 and 4.0 - 7.0. The 2-DE gels held about 300 to 700 detectable protein spots. We randomly picked 12 and nine spots that were consistently expressed in the pH 3.0 - 10.0 and pH 4.0 - 7.0 ranges, respectively. Following MALDI-TOF MS analysis, the 21 randomly selected proteins included proteins known to be present in bovine milk, such as albumin, lactoferrin, serum albumin precursor, T cell receptor, polymeric immunoglobulin receptor, pancreatic trypsin inhibitor, aldehyde oxidase and microglobulin. These proteins have major functions in immune responses, metabolism and protein binding. In summary, we herein identified both known and novel whey proteins present in bovine early milk, and our sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis revealed their expression pattern.

Non-Ionic Surfactants Antagonize Toxicity of Potential Phenolic Endocrine-Disrupting Chemicals, Including Triclosan in Caenorhabditis elegans

  • Alfhili, Mohammad A.;Yoon, Dong Suk;Faten, Taki A.;Francis, Jocelyn A.;Cha, Dong Seok;Zhang, Baohong;Pan, Xiaoping;Lee, Myon-Hee
    • Molecules and Cells
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    • v.41 no.12
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    • pp.1052-1060
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    • 2018
  • Triclosan (TCS) is a phenolic antimicrobial chemical used in consumer products and medical devices. Evidence from in vitro and in vivo animal studies has linked TCS to numerous health problems, including allergic, cardiovascular, and neurodegenerative disease. Using Caenorhabditis elegans as a model system, we here show that short-term TCS treatment ($LC_{50}$: ~0.2 mM) significantly induced mortality in a dose-dependent manner. Notably, TCS-induced mortality was dramatically suppressed by co-treatment with non-ionic surfactants (NISs: e.g., Tween 20, Tween 80, NP-40, and Triton X-100), but not with anionic surfactants (e.g., sodium dodecyl sulfate). To identify the range of compounds susceptible to NIS inhibition, other structurally related chemical compounds were also examined. Of the compounds tested, only the toxicity of phenolic compounds (bisphenol A and benzyl 4-hydroxybenzoic acid) was significantly abrogated by NISs. Mechanistic analyses using TCS revealed that NISs appear to interfere with TCS-mediated mortality by micellar solubilization. Once internalized, the TCS-micelle complex is inefficiently exported in worms lacking PMP-3 (encoding an ATP-binding cassette (ABC) transporter) transmembrane protein, resulting in overt toxicity. Since many EDCs and surfactants are extensively used in commercial products, findings from this study provide valuable insights to devise safer pharmaceutical and nutritional preparations.

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%.

Ginsenoside F4 inhibits platelet aggregation and thrombus formation by dephosphorylation of IP3RI and VASP

  • Shin, Jung-Hae;Kwon, Hyuk-Woo;Lee, Dong-Ha
    • Journal of Applied Biological Chemistry
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    • v.62 no.1
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    • pp.93-100
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    • 2019
  • The root of Panax ginseng is used in ethnomedicine throughout eastern Asia and various recent studies have proved that Panax ginseng has inhibitory effects on cardiovascular disease. Each factor causing cardiovascular disease is known to have a very complex process which is achieved by a diverse number of mechanisms. Among these factors, platelets are the most important because they directly participate in thrombogenesis. Therefore, inhibiting the activity of platelets is an essential element for prevention of cardiovascular diseases. Our previous study showed the antiplatelet effects of Korean red ginseng extract and two of its components, ginsenoside Rg3 and ginsenoside Ro. However, the inhibitory mechanism of other ginsenosides remains unclear. Therefore, we investigated the inhibitory mechanism of ginsenoside F4 (G-F4) from Korean red ginseng on the regulation of signaling molecules involved in human platelet aggregation. With the use of G-F4, collagen-induced human platelet aggregation was inhibited in a dose-dependent manner, and it suppressed collagen-induced elevation of $[Ca^{2+}]_i$ mobilization through elevated phosphorylation of inositol 1, 4, 5-triphosphate receptor I ($Ser^{1756}$). In addition, G-F4 inhibited fibrinogen binding to ${\alpha}IIb/{\beta}_3$ during collagen-induced human platelet aggregation. Thus, in the present study, G-F4 showed an inhibitory effect on human platelet activation, suggesting its potential use as a new natural medicine for preventing platelet-mediated cardiovascular diseases.

Ameliorative Effects of NXP031 on MPTP-Induced Neurotoxicity (MPTP로 유도된 신경 독성에 대한 NXP031의 개선 효과)

  • Lee, Joo Hee;Song, Min Kyung;Kim, Youn-Jung
    • Journal of Korean Biological Nursing Science
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    • v.23 no.3
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    • pp.199-207
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    • 2021
  • Purpose: The purpose of this study was to investigate effects of NXP031, an inhibitor of oxidation by specifically binding to the complex of DNA aptamer/vitamin C, on dopaminergic neurons loss and the reaction of microglia in an animal model of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced subchronic Parkinson's disease (PD). Methods: A subchronic PD mouse model was induced via an intraperitoneal (IP) injection of MPTP 30 mg/kg per day for five days. NXP031 (vitamin C/aptamer at 200 mg/4 mg/kg) and vitamin C at 200 mg/kg were administered via IP injections at one hour after performing MPTP injection. This process was performed for five days. Motor function was then evaluated with pole and rotarod tests, after which an immunohistochemical analysis was performed. Results: NXP031 administration after MPTP injection significantly improved motor functions (via both pole and rotarod tests) compared to the control (MPTP injection only) (p<.001). NXP031 alleviated the loss of dopaminergic neurons in the substantia nigra (SN) and striatum caused by MPTP injection. It was found to have a neuroprotective effect by reducing microglia activity. Conclusion: NXP031 can improve impaired motor function, showing neuroprotective effects on dopaminergic neurons in the SN and striatum of MPTP-induced subchronic Parkinson's disease mouse model. Results of this study suggest that NXP031 has potential in future treatments for PD and interventions for nerve recovery.

Forebrain glutamatergic neuron-specific Ctcf deletion induces reactive microgliosis and astrogliosis with neuronal loss in adult mouse hippocampus

  • Kwak, Ji-Hye;Lee, Kyungmin
    • BMB Reports
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    • v.54 no.6
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    • pp.317-322
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    • 2021
  • CCCTC-binding factor (CTCF), a zinc finger protein, is a transcription factor and regulator of chromatin structure. Forebrain excitatory neuron-specific CTCF deficiency contributes to inflammation via enhanced transcription of inflammation-related genes in the cortex and hippocampus. However, little is known about the long-term effect of CTCF deficiency on postnatal neurons, astrocytes, or microglia in the hippocampus of adult mice. To address this, we knocked out the Ctcf gene in forebrain glutamatergic neurons (Ctcf cKO) by crossing Ctcf-floxed mice with Camk2a-Cre mice and examined the hippocampi of 7.5-10-month-old male mice using immunofluorescence microscopy. We found obvious neuronal cell death and reactive gliosis in the hippocampal cornu ammonis (CA)1 in 7.5-10-month-old cKO mice. Prominent rod-shaped microglia that participate in immune surveillance were observed in the stratum pyramidale and radiatum layer, indicating a potential increase in inflammatory mediators released by hippocampal neurons. Although neuronal loss was not observed in CA3, and dentate gyrus (DG) CTCF depletion induced a significant increase in the number of microglia in the stratum oriens of CA3 and reactive microgliosis and astrogliosis in the molecular layer and hilus of the DG in 7.5-10-month-old cKO mice. These results suggest that long-term Ctcf deletion from forebrain excitatory neurons may contribute to reactive gliosis induced by neuronal damage and consequent neuronal loss in the hippocampal CA1, DG, and CA3 in sequence over 7 months of age.

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.