• Proceedings of the Korean Biophysical Society Conference
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• 1997.07a
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• pp.44-44
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• 1997

Ca$^{2＋}$ is one of the most common metal ions associated with proteins, playing more or less well-defined functional roles in biological activities. In protease, the presence of $Ca^{2＋}$ slows down autolysis and enhances thermal stability. Subtilisin, one of the best studied proteases, is known to have two $Ca^{2＋}$ -binding sites.(omitted)

• Journal of the Korean Magnetic Resonance Society
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• v.19 no.3
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• pp.112-118
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• 2015

Hsp31 protein is one of the members of DJ-1 superfamily proteins and has a dimeric structure of which molecular weight (MW) is 62 kDa. The mutation of DJ-1 is closely related to early onset of Parkinson's disease. Hsp31 displays $Zn^{+2}$-binding activity and was first reported to be a holding chaperone in E. coli. Its additional glyoxalase III active has recently been characterized. Moreover, an incubation at $60^{\circ}C$ induces Hsp31 protein to form a high MW oligomer (HMW) in vitro, which accomplishes an elevated holding chaperone activity. The NMR technique is elegant method to probe any local or global structural change of a protein in responses to environmental stresses (heat, pH, and metal). Although the presence of the backbone chemical shifts (bbCSs) is a prerequisite for detailed NMR analyses of the structural changes, general HSQC-based triple resonance experiments could not be used for 62 kDa Hsp31 protein. Here, we prepared the per-deuterated Hsp31 and performed the TROSY-based triple resonance experiments for the bbCSs assignment. Here, detailed processes of per-deuteration and the NMR experiments are described for other similar NMR approaches.

• Ocean Science Journal
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• v.41 no.1
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• pp.43-51
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• 2006

Concentration and distribution of Fe, Zn, Cu, Cd, Mn, Pb, Ni among subcellular fractions (cellular membrane structures and cytosol) and Zn, Cu, Cd among cytoplasmic proteins in the kidney and digestive gland of mussel Modiolus modiolus living along a polymetallic concentration gradient were studied. It was found in the kidney of M. modiolus from contaminated sites that the Fe percent increased in the "membrane" fraction, whereas Zn, Pb, Ni and Mn percent increased in the cytosol compared to the kidney of the control mussel. Note kidney cytosol of M. modiolus from clean and contaminated sites sequestered major parts of Cu and Cd. In the digestive gland of M. modiolus from contaminated sites Fe, Zn, Cd, Mn, Ni percent increased in the "membrane" fraction, whereas Cu, Pb percent increased in the cytosol compared to digestive gland of control mussel. Gel-filtration chromatography shows kidney of M. modiolus contains increased metallothionein-like protein levels irrespective of ambient dissolved metal concentrations. It was shown that the metal detoxification system in the kidney and digestive gland of M. modiolus was efficient under extremely high ambient metal levels. However, under complex environmental contamination in the kidney of M. modiolus, the metal detoxification capacity of metallothionein-like proteins was damaged.

• Korean Journal of Microbiology
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• v.50 no.1
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• pp.73-83
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• 2014

The objective of the this study was to investigate the binding capacity and removal ability of lactic acid bacterial strains obtained from Korean soybean paste for mutagenic heterocyclic amines (HCAs) formed during cooking of protein-rich food at high temperature. Among 19 strains identified by carbohydrate fermentation and 16S rRNA sequencing, the live cell or cell-free culture supernatant of Lactobacillus acidophilus D11, Enterococcus faecium D12, Pediococcus acidilactici D19, L. acidophilus D38, Lactobacillus sakei D44, Enterococcus faecalis D66, and Lactobacillus plantarum D70 inhibited the mutagenesis caused by either 3-amino-1,4-dimethyl-5H-pyrido[4,3-b] indole (Trp-P-1) or 3-amino-1-methyl-5H-pyrido[4,3-b] indole (Trp-P-2) in Salmonella typhimurium TA98 and TA100. The bacterial cells of the isolated strains showed greater binding activity than the pure cell wall, exopolysaccharide, and pepetidoglycan. The carbohydrate moieties of the cell wall or protein molecules on the cell surface have a significant role in binding Trp-P-1 and Trp-P-2, since protease, heating, sodium metaperiodate, or acidic pH treatments significantly (P<0.05) reduced the binding efficacy of the tested bacteria. Addition of metal ions or sodium dodecyl sulfate decreased the binding ability of E. faecium D12, L. acidophilus D38, and E. faecalis D66. Therefore, the binding mechanisms of these strains may consist of ion-exchange and hydrophobic bonds. Especially, the high mutagen binding by L. acidophilus D38 and L. plantarum D70 may reduce the accumulation or absorption of Trp-P-1 and Trp-P-2 in the small intestine via increased excretion of a mutagen-bacteria complex.

• The Korean Journal of Ecology
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• v.24 no.1
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• pp.35-43
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• 2001

Accumulation, elimination and subcellular distribution of heavy metals in Littorina brevicula exposed to cadmium and zinc separately and concurrently were investigated. When the winkles had been exposed to 400 ㎍/L CdCl₂ and 3000 ㎍/l ZnSO₄ separately for 90 days, each of the metal body burden in the whole sofl parts increased in proportion to time of exposure until 70 days. But it didn't increase after 70 days. But when the winkles had been exposed to cadmium and zinc simultaneously, cadmium body burden decreased but zinc body burden increased as compared to the winkles exposed to each of the metal. We also found that cadmium accumulated in the winkles was not depurated for 42 days, but zinc accumulated in them was depurated. Especially, zinc was depurated faster when they had been exposed to mixture of cadmium and zinc. After the winkles had been exposed to cadmium and zinc separately for 70 days, about 60% cadmium of the total body burden was associated with the soluble fraction, while about 75% zinc of the total body burden was associated with insoluble fraction. And these trends of metal partitioning did not alter when the winkles had been exposed to metal mixture. After the soluble fraction applied to gel-filtration chromatography column, the distribution patterns of cadmium and zinc associated with proteins or ligands were different each other. Most of cadmium (＞90%) in the soluble fraction was bound to MBP-1 (Metal-binding protein-1), about 6.5 kDa), while zinc was distributed evenly to HMW (High molecular weight fraction, >60 kDa), MBP-1, MBP-2 (about 5 kDa), LMW (Low molecular weight fraction, <1 kDa).

• The Korean Journal of Ecology
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• v.22 no.2
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• pp.95-100
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• 1999

The heavy metal concentrations in seawater and winkles (Littorina brevicula) collected from the Onsan bay area in southeast of Korea were analyzed. The heavy metal concentrations in the seawater obtained from the most polluted site showed approximately 189. 205. and 110 fold higher cadmium, copper. and zinc concentrations, respectively. than the uncontaminated control site. The contamination levels of these metals in winkles were 11.08 - 2.35, 334.5 - 212.5, and 426.0 - 499.2$\mu\textrm{g}$ per gram dry body weight. respectively. The concentrations of all three metals in both the seawater and winkles decreased gradually with increasing distance from Daejeong stream, suggesting the stream being the major source of heavy metal input into the bay. Among the four body parts of digestive gland and gonad. gill. kidney, and remaining tissue in contaminated winkles, kidney showed the highest accumulation level of cadmium: copper and zinc, however. were more or less distributed among the four body parts. Upon gel filtration chromatography of the cytosol from the kidney of cadmium induced winkles, one cadmium peak corresponded to the elution peak of horse kidney metallothionein.

• Journal of Periodontal and Implant Science
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• v.38 no.sup2
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• pp.343-354
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• 2008

Purpose: Aggregatibacter actinomycetemcomitans is associated with localized aggressive periodontitis. It produces cytolethal distending toxin (CDT), which induces cell cycle arrest in the G2/M phase. The CDT holotoxin is composed of CdtA, CdtB, and CdtC. CdtB has structural homology to human DNase I and is an active component of the CDT complex acting as a DNase. In particular, the pattern homology seen in the CdtB subunit has been associated with specific DNase I residues involved in enzyme catalysis, DNA binding, and metal ion binding. So, to study the functions and regulation of recombinant CdtB, we made up a quantity of functional recombinant CdtB and tested it in relation to the metal ion effect. Materials and Methods: We constructed the pET28a-cdtB plasmid from A. actinomycetemcomitans Y4 by genomic DNA PCR and expressed it in the BL21 (DE3) Escherichia coli system. We obtained the functional recombinant CdtB by the refolding system using the dialysis method and then analyzed the DNase activity and investigated the metal ion effect from plasmid digestion. Results: The recombinant CdtB subunit was expressed as the inclusion bodies. We were able to obtain functional recombinant CdtB subunit using refolding system. We confirmed that our refolded recombinant CdtB had DNase activity and was influenced by the metal ions $Mg^{2+}$ and $Ca^{2+}$. Conclusion: We suggest that the factors influencing recombinant CdtB may contribute to CDT associated diseases, such as periodontitis, endocarditic, meningitis, and osteomyelitis.

• Journal of Microbiology and Biotechnology
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• v.31 no.2
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• pp.233-239
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• 2021

Cyanobacteriochromes (CBCRs) are phytochrome-related photoreceptor proteins in cyanobacteria and cover a wide spectral range from ultraviolet to far-red. A single GAF domain that they contain can bind bilin(s) autocatalytically via heterologous recombination and then fluoresce, with potential applications as biomarkers and biosensors. Here, we report that a novel red/green CBCR GAF domain, SPI1085g2 from Spirulina subsalsa, covalently binds both phycocyanobilin (PCB) and phycoerythrobilin (PEB). The PCB-binding GAF domain exhibited canonical red/green photoconversion with weak fluorescence emission. However, the PEB-binding GAF domain, SPI1085g2-PEB, exhibited an intense orange fluorescence (λabs.max = 520 nm, λfluor.max = 555 nm), with a fluorescence quantum yield close to 1.0. The fluorescence of SPI1085g2-PEB was selectively and instantaneously quenched by copper ions in a concentration-dependent manner and exhibited reversibility upon treatment with the metal chelator EDTA. This study identified a novel PEB-binding cyanobacteriochrome-based fluorescent protein with the highest quantum yield reported to date and suggests its potential as a biosensor for the rapid detection of copper ions.

• Journal of Animal Science and Technology
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• v.63 no.5
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• pp.1159-1168
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• 2021

Ovotransferrin (OTF), an egg protein known as transferrin family protein, possess strong antimicrobial and antioxidant activity. This is because OTF has two iron binding sites, so it has a strong metal chelating ability. The present study aimed to evaluate the improved immune-enhancing activities of OTF hydrolysates produced using bromelain, pancreatin, and papain. The effects of OTF hydrolysates on the production and secretion of pro-inflammatory mediators in RAW 264.7 macrophages were confirmed. The production of nitric oxide (NO) was evaluated using Griess reagent and the expression of inducible nitric oxide synthase (iNOS) were evaluated using quantitative real-time polymerase chain reaction (PCR). And the production of pro-inflammatory cytokines (tumor necrosis factor [TNF]-α and interleukin [IL]-6) and the phagocytic activity of macrophages were evaluated using an ELISA assay and neutral red uptake assay, respectively. All OTF hydrolysates enhanced NO production by increasing iNOS mRNA expression. Treating RAW 264.7 macrophages with OTF hydrolysates increased the production of pro-inflammatory cytokines and the phagocytic activity. The production of NO and pro-inflammatory cytokines induced by OTF hydrolysates was inhibited by the addition of specific mitogen-activated protein kinase (MAPK) inhibitors. In conclusion, results indicated that all OTF hydrolysates activated RAW 264.7 macrophages by activating MAPK signaling pathway.

• Proceedings of the Materials Research Society of Korea Conference
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• 2011.05a
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• pp.10.2-10.2
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• 2011

The design and synthesis of protein-like polymers is a fundamental challenge in materials science. A biomimetic approach is to explore the impact of monomer sequence on non-natural polymer structure and function. We present the aqueous self-assembly of two peptoid polymers into extremely thin two-dimensional (2D) crystalline sheets directed by periodic amphiphilicity, electrostatic recognition and aromatic interactions. Peptoids are sequence-specific, oligo-N-substituted glycine polymers designed to mimic the structure and functionality of proteins. Mixing a 1:1 ratio of two oppositely charged peptoid 36 mers of a specific sequence in aqueous solution results in the formation of giant, free-floating sheets with only 2.7 nm thickness. Direct visualization of aligned individual peptoid chains in the sheet structure was achieved using aberration-corrected transmission electron microscopy. Specific binding of a protein to ligand-functionalized sheets was also demonstrated. The synthetic flexibility and biocompatibility of peptoids provide a flexible and robust platform for integrating functionality into defined 2D nanostructures. In the later part of my talk, we describe the use of metal ions to construct two-dimensional hybrid films that have the ability to self-heal. Incubation of biomimetic peptoid polymers with specific divalent metal ions results in the spontaneous formation of uniform multilayers at the air-water interface. We anticipate that ease of synthesis and transfer of these two-dimensional materials may have many potential applications in catalysis, gas storage and sensing, optics, nanomaterial synthesis, and environmentally responsive scaffolds.