• Bulletin of the Korean Chemical Society
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• v.24 no.6
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• pp.817-823
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• 2003

We have performed ab initio calculation on the active site of HIV-1 protease. The FEP method was used to determine the binding free energy of four different of protonated states of HIV-1 protease with inhibitor. The structure of the active site and hole structure was taken from the X-ray crystallographic coordinates of the C₂ symmetric inhibitor A74704 protease bound. The active site was modeled with the fragment molecules of binding pocket, acetic acid/ acetate anion (Asp25, Asp125), formamide (amide bond of Thr26/Gly27, Thr126/ Gly127), and methanol as inhibitor fragment. All possibly protonated states of the active site were considered, which were diprotonated state (0, 0), monoprotonated (-1, 0),(0, -1) and diunprotonated state (-1, -1). Once the binding energy Debind, of each model was calculated, more probabilistic protonated states can be proposed from binding energy. From ab-initio results, the FEP simulations were performed for the three following mutations: Ⅰ) Asp25 … Asp125 → AspH25 … Asp125, ⅱ) Asp25 … Asp125 → Asp25 … AspH125, ⅲ) AspH25 … Asp125 → AspH25 … AspH125. The free energy difference between the four states gives the information of the more realistic protonated state of active site aspartic acid. These results provide a theoretical prediction of the protonation state of the catalytic aspartic residues for A74707 complex, and may be useful for the evaluation of potential therapeutic targets.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.58 no.1
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• pp.50-56
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• 2013

For accurate analysis of low molecular peptides using SELDI-TOF MS (surface enhanced laser desorption/ionization time of flight mass spectrometry), the optimized analytical conditions should be established for a specific biological sample. This study was conducted to optimize SELDI-TOF MS analytical conditions for profiling low molecular peptide below 10 kDa presented in sorghum seeds. Analytical conditions were as follows; (1) protein chips: CM10 (weak cation exchanger) and Q10 (strong anion exchanger), (2) dilution factors of binding buffer: 1/2, 1/5, 1/10, 1/20, 1/50, 1/100, and 1/200, (3) the stringency of Q10 binding buffer: 10 mM and 100 mM, and (4) protein extraction buffers: sodium borate, sodium borate + acetone, phenol, and TCA buffers. Optimum dilution factors were selected as 1/20 and 1/50 in both protein chips, CM10 and Q10. Low stringency of Q10 binding buffer (10mM) detected more peptide peaks than high stringency (100 mM). Selected protein extraction buffers of sorghum seed for SELDI-TOF MS analysis was the sodium borate buffer in the range of 2~10 kDa, while the phenol buffer was more suitable in the range of 10~20 kDa.

• The Korean Journal of Ecology
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• v.21 no.3
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• pp.225-231
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• 1998

The studies on the potentiality of biomonitoring heavy metal pollution in coastal region of industrial complex were performed to investigate the heavy metal accumulation and induction of metal-binding protein (MBP) as detoxification process using Rumex maritimus. Bioconcentration in organs and MBP in root of R. maritimus was investigated for the research of the tolerance of heavy metals. The bioconcentration of cadmium and zinc in organs showed 3.6-8.0 times in root higher than in shoot, so it was found that heavy metal accumulated selectively in root. MBP increased absorbance in 254 nm and decreased in 280 nm, because it was composed of high cystein content and low aromatic acids, so absorbance had large difference between 254 nm and 280 nm. The existence of MBP in the 10-20 fraction was ascertained with anion exchange chromatography and it was identified that concentration of heavy metal increased according as an exposure concentration of medium increased in QAE Sephadex A-25 elution profile. These results suggested that MBP could play a role in biomarker determining the bioconcentration of plant. This study demonstrated a possibility that removal ability of heavy metal of R. maritimus resulted from detoxification process and MBP could be utilized as a biomarker of heavy metal pollution.

• Bulletin of the Korean Chemical Society
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• v.17 no.12
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• pp.1127-1131
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• 1996

[FeⅡ2(N-Et-HPTB)Cl2]BPh4(1), where N-Et-HPTB is the anion of N,N,N',N'-tetrakis(N-ethyl-2-benzimidazolylmethyl)-2-hydroxy-l,3-diaminopropane, has been synthesized to model dioxygen binding to the diferrous centers of proteins. 1 has a singly bridged structure with a μ-alkoxo of N-Et-HPTB and contains two five-coordinate iron(Ⅱ) centers with two chloride ligands as exogenous ligands. 1 exhibits an electronic spectrum with a λmax at 336 nm in acetone. 1 in acetone exhibits no EPR signal at 4 K, indicating diiron(Ⅱ) centers are antiferromagnetically coupled. Exposure of acetone solution of 1 to O2 at -90 ℃ affords an intense blue color intermediate showing a broad band at 586 nm. This absorption maximum of the dioxygen adduct(1/O2) was found in the same region of μ-l,2-peroxo diiron(Ⅲ) intermediates in the related complexes with pendant pyridine or benzimidazole ligand systems. However, this blue intermediate exhibits EPR signals at g = 1.93, 1.76, and 1.59 at 4 K. These g values are characteristic of S = 1/2 system derived from an antiferromagnetically coupled high-spin Fe(Ⅱ)Fe(Ⅲ) units. 1 is the unique example of a (μ-alkoxo)diferrous complex which can bind dioxygen and form a metastable mixed-valence intermediate. At ambient temperature, most of 1/O2 intermediate decays to form a diamagnetic species. It suggests that the dacay reaction of the intermediate might be bimolecular, implying the formation of mixed-valence tetranuclear species in transition state.

• KSBB Journal
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• v.9 no.2
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• pp.98-103
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• 1994

A bed configuration wherein sheets of modified fibrous polyethylene are potted within a Millipore Filter Cartridge matrix has been developed. Polyethylene fibers form sturdy beds but the native hydrophobicity and inertness of polyethylene have precluded their use in protein chromatography The polyethylene fibers used in this system were modified by plasma oxidation and further derivatization. The resulting fibers are hydrophilic, bind protein reversibly and serve as an anion-exchange stationary phase. Separation of Bovine Serum Albumin on this bed, as well as results of basic studies on capacity and reversibility of binding within a fibrous bed and experimental data handling system are shown.

• Molecules and Cells
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• v.38 no.8
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• pp.741-741
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• 2015

• Journal of Microbiology and Biotechnology
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• v.19 no.7
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• pp.713-717
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• 2009

The dependence of the catalytic properties of horseradish peroxidase on the structural changes of ionic liquids was investigated with two water-miscible ionic liquids, N-butyl-3methypyridinium tetraftuoroborate ([$BMP_y$][$BF_4$]) and 1-butyl-3-methylimidazolium methylsulfate ([BMIM][$MeSO_4$]), each of which shares an anion ($BF_4^-$) or a cation ($BMIM^+$) with 1-butyl-3-methylimidazolium tetraftuoroborate ([BMIM][$BF_4$]), respectively. The oxidation of guaiacol (2-methoxyphenol) with $H_2O_2$was used as a model reaction. In order to minimize the effect of solution viscosity on the kinetic constants of the enzymatic catalysis, the enzymatic reactions for the kinetic study were performed in water-ionic liquid mixtures containing 25% (v/v) ionic liquid at maximum. Similarly to the previously reported results for [BMIM][$BF_4$], as the concentration of [$BMP_y$][$BF_4$] increased, the $K_m$value increased with a decrease in the $k_{cat}$value: the $K_m$value increased markedly from 2.8 mM in 100% water to 12.6 mM in 25% (v/v) ionic liquid, indicating that ionic liquid significantly weakens the binding affinity of guaiacol to the enzyme. On the contrary, [BMIM][$MeSO_4$] decreased the Km value to 1.4 mM in 25% (v/v) ionic liquid. [BMIM][$MeSO_4$] also decreased $k_{cat}$more than 3-folds [from 13.8 $s^{-1}$in 100% water to 4.1 $s^{-1}$in 25% (v/v) ionic liquid]. These results indicate that the ionic liquids interact with the enzyme at the molecular level as well as at a macroscopic thermodynamic scale. Specifically, the anionic component of the ionic liquids influenced the catalysis of horseradish peroxidase in different ways.

• Macromolecular Research
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• v.12 no.1
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• pp.46-52
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• 2004

In a continuation of our previous studies on blood compatibility profiles of anion-substituted poly(vinyl alcohol) (PVA) membranes, in which hydroxyl groups have been replaced with carboxymethyl (C-PVA) and sulfonyl groups (S-PVA), we have studied the activation of complement components and the changes in white cell and platelet count in vitro and compared them with those of unmodified PVA, Cuprophane, and low-density polyethylene. Complement activation of fluid phase components, C3a, Bb, iC3b, and SC5b-9, and of bound phases, C3c, C3d, and SC5b-9, were assessed by enzyme-linked immunosorbent assay (ELISA) and immunoblot, respectively. The changes in the number of white cells and platelets following complement activation were counted using a Coulter counter. C-PVA and S-PVA activated C3 to a lesser extent than did PVA, which we attribute to the diminished level of surface nucleophiles of the samples. In addition, C- and S-PVA exhibit increased inhibition of Bb production, resulting in a decrease in the extent of C5 activation. Consequently, because of the reduced activation of C3 and C5, C- and S-PVA samples cause marked decreases in the SC5b-9 levels in plasma. We also found that the negatively charged sulfonate and carboxylate groups of the samples cause a greater extent of adsorbtion of the positively charged anaphylatoxins, C3a and C5a, because of strong electrostatic attraction, which in turn provides an inhibition of chemotaxis and activation of leukocytes. The ability to inhibit complement production, together with the binding ability of anaphylatoxins of the C- and S-PVA samples, leads to a prominent decrease in lysis of leukocytes as well as activation of platelets.

• Applied Chemistry for Engineering
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• v.9 no.3
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• pp.377-382
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• 1998

This study was carried out to investigate the relation between surface acidity and interfacial electrical characteristics of surface-treated ${\gamma}-alumina$ whose surface activity was increased. The points of zero charge (P. Z. C.) of ${\gamma}-alumina$ whose surface is treated with the sulfuric, nitric and hydro-chloric acid of various concentration were measured from the site-binding theory and mass transport method. The surface active sites were measured by amine titration method and Hammett indicator method. The interfacial properties at alumina/KCl(aq) interface were measured by potentiometric titration. From the experimental results, the following results were obtained. Pure ${\gamma}-alumina$ surface acidity decreases with the increase of calcination temperature at strength $H_o{\leq}+9.3$ Surface-treated alumina acidic properties increase with the anion loading on alumina surface. The surface ionization constants decrease with anion loading on alumina surface, then P. Z. C. decreases with acid amount on alumina surface. Acid amount of surface treated alumina can be correlated with surface charge density at strength $H_o{\leq}+4.8$ as follows. $SO_4^2-/Al_2O_3:Q_A=-0.172ln(0.0418{\sigma}+1.448)$ $NO_3^-/Al_2O_3:Q_A=-0.024{\sigma}-0.0189$ $Cl^-/Al_2O_3:Q_A=-0.01{\sigma}-0.2006$.

• Asian-Australasian Journal of Animal Sciences
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• v.30 no.5
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• pp.643-652
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• 2017

Objective: Prostaglandins (PGs) function in various reproductive processes, including luteolysis, maternal pregnancy recognition, conceptus development, and parturition. Our earlier study has shown that PG transporters ATP-binding cassette, subfamily C, member 4 (ABCC4) and solute carrier organic anion transporter family, member 2A1 (SLCO2A1) are expressed in the uterine endometrium in pigs. Since several other PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 are known to be present in the uterine endometrium, this study investigated the expression of these PG transporters in the porcine uterine endometrium and placenta. Methods: Uterine endometrial tissues were obtained from gilts on day (D) 12 and D15 of the estrous cycle and days 12, 15, 30, 60, 90, and 114 of pregnancy. Results: ABCC1, ABCC9, SLCO4C1, and SLCO5A1 mRNAs were expressed in the uterine endometrium, and levels of expression changed during the estrous cycle and pregnancy. Expression of ABCC1 and ABCC9 mRNAs was localized mainly to luminal and glandular epithelial cells in the uterine endometrium, and chorionic epithelial cells during pregnancy. Conceptuses during early pregnancy and chorioallantoic tissues from mid to late pregnancy also expressed these PG transporters. $Estradiol-17{\beta}$ increased the expression of ABCC1 and SLCO5A1, but not ABCC9 and SLCO4C1 mRNAs and increasing doses of $interleukin-1{\beta}$ induced the expression of ABCC9, SLCO4C1, and SLCO5A1 mRNAs in endometrial explant tissues. Conclusion: These data showed that several PG transporters such as ABCC1, ABCC9, SLCO4C1, and SLCO5A1 were expressed at the maternal-conceptus interface, suggesting that these PG transporters may play an important role in the establishment and maintenance of pregnancy by regulating PG transport in the uterine endometrium and placenta in pigs.