• Title/Summary/Keyword: Anion binding

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Studies on the Separation of Immunoglobulin and Immunological Response from Korean Native Cattle (한우초유중 Ig의 분리 ${\cdot}$ 정제 및 면역 반응에 관한 연구)

  • Baick, Seung-Cheon;Kim, Yong-Hwi;Shin, Je-Ho;Yu, Je-Hyeon
    • Journal of Dairy Science and Biotechnology
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    • v.15 no.1
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    • pp.1-9
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    • 1997
  • This study was conducted to efficiently separate the Ig from Korean native cattle colostrum and to utilize them as an immunogen for the production of antibodies aginst rabbit. The results obtained were as follows : 1. About 84% of Ig G could be separated from Korean native cattle colostrum by·gel filtration using Superose 12 column on HPLC. The separation profile of Korean native cattle colostral immunoglobulin was similar that of Holstein colostral Ig. 2. Separation of Korean native cattle colostral Ig by anion exchange chromatography using Mono Q column on HPLC was poor resolution chromatographic pattern. 3. Hi-Trap Protein G column showed better results than the Protein A Sepharose CL-4B column in the Ig G binding capacity from Korean native cattle colostral Ig. 4. Protein G Sepharose Fast Flow system resulted in higher Ig g binding capacity as the industrial size scale-up approach. 5. Sufficient titer reaction of antibody to Korean native cattle colostral Ig G was confirmed by ELISA.

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Biodistribution of 99mTc Labeled Integrin Antagonist

  • Jang, Beom-Su;Park, Seung-Hee;Shin, In Soo;Maeng, Jin-Soo;Paik, Chang H.
    • Toxicological Research
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    • v.29 no.1
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    • pp.21-25
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    • 2013
  • The selective targeting of an integrin ${\alpha}_v{\beta}_3$ receptor using radioligands may enable the assessment of angiogenesis and integrin ${\alpha}_v{\beta}_3$ receptor status in tumors. The aim of this research was to label a peptidomimetic integrin ${\alpha}_v{\beta}_3$ antagonist (PIA) with $^{99m}Tc(CO)_3$ and to test its receptor targeting properties in nude mice bearing receptor-positive tumors. PIA was reacted with tris-succinimidyl aminotriacetate (TSAT) (20 mM) as a PIA per TSAT. The product, PIA-aminodiacetic acid (ADA), was radiolabeled with $[^{99m}Tc(CO)_3(H_2O)_3]^{+1}$, and purified sequentially on a Sep-Pak C-18 cartridge followed by a Sep-Pak QMA anion exchange cartridge. Using gradient C-18 reverse-phase HPLC, the radiochemical purity of $^{99m}Tc(CO)_3$-ADA-PIA (retention time, 10.5 min) was confirmed to be > 95%. Biodistribution analysis was performed in nude mice (n = 5 per time point) bearing receptor-positive M21 human melanoma xenografts. The mice were administered $^{99m}Tc(CO)_3$-ADA-PIA intravenously. The animals were euthanized at 0.33, 1, and 2 hr after injection for the biodistribution study. A separate group of mice were also co-injected with 200 ${\mu}g$ of PIA and euthanized at 1 hr to quantify tumor uptake. $^{99m}Tc(CO)_3$-ADA-PIA was stable in phosphate buffer for 21 hr, but at 3 and 6 hr, 7.9 and 11.5% of the radioactivity was lost as histidine, respectively. In tumor bearing mice, $^{99m}Tc(CO)_3$-ADA-PIA accumulated rapidly in a receptor-positive tumor with a peak uptake at 20 min, and rapid clearance from blood occurring primarily through the hepatobiliary system. At 20 min, the tumor-to-blood ratio was 1.8. At 1 hr, the tumor uptake was 0.47% injected dose (ID)/g, but decreased to 0.12% ID/g when co-injected with an excess amount of PIA, indicating that accumulation was receptor mediated. These results demonstrate successful $^{99m}TC$ labeling of a peptidomimetic integrin antagonist that accumulated in a tumor via receptor-specific binding. However, tumor uptake was very low because of low blood concentrations that likely resulted from rapid uptake of the agent into the hepatobiliary system. This study suggests that for $^{99m}Tc(CO)_3$-ADA-PIA to be useful as a tumor detection agent, it will be necessary to improve receptor binding affinity and increase the hydrophilicity of the product to minimize rapid hepatobiliary uptake.

Kinetic and Chemical Mechanism of Pig Liver Methenyltetrahydrofolate Synthetase (돼지 간 유래의 methenyltetrahydrofolate synthetase 반응메커니즘)

  • Cho, Yong-Kweon
    • Journal of Life Science
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    • v.18 no.8
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    • pp.1036-1041
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    • 2008
  • Methenyltetrahydrofolate synthetase extract was obtained from mouse liver and purified via $30{\sim}70%$ ammonium sulfate fractionation, Fast Q anion exchange and phenyl agarose chromatography. HPLC gel chromatography and SDS-polyacrylamide electrophoresis experiments showed that the enzyme is a monomer with molecular weight of 23 kDa. Optimum temperature and pH were $35^{\circ}C$ and 6.5, respectively. The enzyme was chemically modified only by tetranitromethane and 1-ethyl-3- (3-dimethyl aminopropyl)-carbodiimide (EDC), indicating that tyrosine and carboxylate are in the active site. pH studies showed that 2 tyrosines are involved in the binding of the substrates and a carboxylate in catalysis. Therefore, the chemical mechanism of the enzyme is likely that 2 tyrosines bind to ATP and 5-formylTHFand a carboxylate acts as a general base.

A Study on Ion Distribution and Behavior of Acidrain in Mokpo and Yeochon Area (목포, 여천지역 강수의 무기이온 성분농도와 거동에 관한 연구)

  • 오길영;양수인;이완진
    • Journal of Korean Society for Atmospheric Environment
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    • v.15 no.4
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    • pp.385-392
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    • 1999
  • This study was carried out to investigate the constituents of inorganic ions of precipitation in Mokpo and Yeochon area from Jan. 1996 to Dec. 1997. The volume-weighted mean pH of precipitation was 5.7(4.8~7.8) at Mokpo, and 5.6(5.1~7.4) at Yeochon area, respectively. The non-seasalt(nss) anion concentrations were found in order of $nss-SO^{2-}_4>NO^-_3>nss-Cl^-$ at two areas. The portion of $nss-SO^{2-}_4;and;NO^-_3$ was 71~84% out of anions. The the non-seasalt cation concentrations were found in order of $NH^+_4>nss-Ca^{2-}>nss-Mg^{2+}>nss-K^-$ at two areas. The portion of $NH^+_4;and; nss-Ca^{2+}$ was 85~92% out cations. Compared regionally the year concentraton of $nss-SO^{2-}_4$, the result of Yeochon was 3 times higher than that of Mokpo in 1996, while the results of two regions were almost similar in 1997. The reason was that Yeochon was restricted area of the use of 0.5% sulfur B-C oil, controlled air pollution emission area, controlled semitotal amounts of air pollution emission area and partial operating of factories in 1997. There were no ions having high correlation with $H^+,;but;nss-SO^{2-}_4,;NO^-_3,;NH^+_4;and;nss-Ca^{2+}$ showed high correlation coefficient each other. It seems that these ions have little correlation with $H^+$ because they are washed out on binding state. Factor analysis showed that the first factor was complicated factor containing anthropogenic and soil resource, the second factor was sea-salt resource and the third factor was independent behavior of hydrogen at Mokpo. While, the first factor was complicated factor containing anthropogenic and sea-salt resource, the second factor was $Ca(NO_3)_2$ salt and the third factor was $NH^+_4;and;SO^{2-}_4$ synergied by resource and combination at Yeochon.

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Bone-like Apatite Formation on Ultrafine-Structure in Modified Electrolytic Solution

  • Jang, Jae-Myung;Choe, Han-Cheol
    • Proceedings of the Korean Institute of Surface Engineering Conference
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    • 2017.05a
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    • pp.155-155
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    • 2017
  • Surface modifications are commonly utilized to adjust the properties of the titanium and its alloy surface to the specific needs of the medical applications, but there are disadvantages such as poor osteoconductive properties and low adhesion of bone cell to implant surface. In order to improve these disadvantages, changes in surface properties have an important effect on osseointegration during implantation. In this paper we applied new technological method for improving a unique surface modification using the characteristic of an electrolytic Solution. Thus, in the electrolyte containing NaF in Na2SO4, TiO2 nanoporous was uniformly formed, and HAp nanoparticles were electrodeposited around the TiO2 nanopores, but in the electrolyte containing NH4F in (NH4)H2PO4, the coarse protrusions including HAp nano particles were regularly deposited onto the TiO2 barrier layer. The surface characteristics and the distributed elements and have been investigated by EDS analysis, and ultra-fine structure of surface are carried out using FE-SEM. To investigate the behavior of the anion, the analysis of chemical states was performed by XPS, and the narrow spectrums for Ti2P, Ca 2p, and P 2p seems to be almost similar depending on the characteristics of the electrolyte solution respectively. In addition, Ca 2p spectrum could be resolved into two peaks for Ca 2p3/2 and 2p1/2 at 347.4 and 351.3 eV, which are related to hydroxyapatite. And, the P peak can also be deconvoluted into two peaks for P1/2 and P3/2 levels with binding energy 134.2 and 133.4 eV, respectively. From the result of soaking test, the apatite morphologys were well-formed onto the modified surface according to the different conditions.

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Characterization and Partial Purification of Storage Protein-i Receptor in the Fat Body of Hyphantria cunea (미국흰불나방 지방체에서 저장단백질-1의 수용체의 특성과 부분정제)

  • 채권석;여성문;김학렬
    • The Korean Journal of Zoology
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    • v.38 no.4
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    • pp.490-497
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    • 1995
  • In vitro tissue culture of fat body of Hyphantria cunea in the medium containing [35S]-methionine reveaied that storage protein-i (SP-1) is taken up into fat body of prepupae and 1-day-old pupae. Using Western blotting and ligand binding method, we were able to identify the protein band of the SP-1 receptor protein. For the partial purification, the membrane proteins of fat body cells were solubilixed with 1% Triton X-1OO and applied to anion exchange chromatography. The results revealed the molecular weight of the receptor protein to be about 80 kl)a in SDSPAGE, and the P1 was estimated to be about 6.1. The mobility of the receptor protein in 8D8-PAGE was highly dependent on both temperature during electrophoresls and the condition of samples whether they were in reducing or nonreducing.

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Acrylic Polymer Composition Suitable for Ion Delivery and Selective Detection of Proton, and Hydroxyl and Cu(II) Ions (이온 이동에 적합한 아크릴고분자 박막의 조성과 수소이온, 수산화이온, 구리이온의 선택적 검출)

  • Lee, Dahye;Woo, Heejung;Do, Jung Yun
    • Polymer(Korea)
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    • v.38 no.6
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    • pp.801-808
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    • 2014
  • Ion diffusion and chemical binding to acrylic polymer were investigated in a solid film. The composition of acrylic monomers containing amino group and carboxylic acid was adjusted for rapid ion migration in the film. p-Methylred (PMR) and phenolphthalein derivatives were optically sensitive to the concentration of proton and hydroxyl anion, respectively and verified the ion migration through the film layers. A rapid proton migration was observed in the film of a high amino content. On the other hand, $OH^-$ migration occurred rapidly in a high content of carboxylic acid group. The proton migration occurred through the internal layer as well as surface layer of a film and was reversible during 50 repetition examination. Copper(II) ion migration was examined with a Rhodamine-containing polymer film. The light absorption and emission spectra of a Rhodamine-Cu complex showed the key contribution of carboxylic acid group to the Cu(II) migration in a film.

Synthesis, Characterization, and Crystal Structures of Iron(Ⅱ) and Manganese(II) Complexes with 4,7-bis(2-pyridylmethyl)-1-thia-4,7-diazacyclononane

  • Delong Zhang;Daryle H. Busch;Nathaniel W. Alcock
    • Bulletin of the Korean Chemical Society
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    • v.19 no.9
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    • pp.897-906
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    • 1998
  • A new synthesis has been developed for 1-thia-4,7-diazacyclononane and the complexation behavior of a particular derivative has been explored. The pentadentate ligand 4,7-bis(2-pyridylmethyl)-l-thia-4,7-diazacyclononane ([9]$N_2SPY_2$) and its iron(Ⅱ) and manganese(Ⅱ) complexes were prepared and characterized. Magnetic moments of 5.17 and 5.90 μB respectively, indicate that the iron(Ⅱ) and manganese(Ⅱ) complexes are high spin. Charge transfer transitions (d-π*) occur for [Fe(Ⅱ)([9]$N_2SPY_2)(X)]^{n+}$at 27027, 25000, and 24390 cm-1 for X=$H_2O$, Cl-, and OH-, respectively. In acetonitrile solution, the cyclic voltammogram of the manganese(Ⅱ) complex exhibits a redox couple at 0.92 V vs. NHE while the redox potentials for [Fe(Il)([9]$N_2SPY_2)(X)]^{n+}$ are 0.70, 0.66, and 0.37 V vs. NHE for X=$H_2O$, Cl-, and OH-, respectively. The d-π* charge transfer energy and Fe(Ⅱ)/Fe(Ⅲ) redox potential for [Fe(Ⅱ)([9]$N_2SPY_2)(X)]^{n+}$ increase in the same order: $H_2O>Cl^- >OH^-$. The crystal structures of the iron(Ⅱ) and manganese(Ⅱ) complexes reveal that the metal ions are sixcoordinate, binding to four nitrogen atoms and a sulfur atom from the pentadentate ligand, as well as a chloride anion, with the chloride and sulfur atoms in cis positions. The two metals have similar coordination geometries, which are closer to trigonal prismatic than octahedral. In both iron and manganese complexes, the M-N($sp_3$) trans to Cl- is 0.07 Å longer than the one cis to Cl- , and M-N($sp^2$) trans to S is 0.05 longer than the one cis to S atom.

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.

Crystal Structure of a Highly Thermostable α-Carbonic Anhydrase from Persephonella marina EX-H1

  • Kim, Subin;Sung, Jongmin;Yeon, Jungyoon;Choi, Seung Hun;Jin, Mi Sun
    • Molecules and Cells
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    • v.42 no.6
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    • pp.460-469
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    • 2019
  • Bacterial ${\alpha}-type$ carbonic anhydrase (${\alpha}-CA$) is a zinc metalloenzyme that catalyzes the reversible and extremely rapid interconversion of carbon dioxide to bicarbonate. In this study, we report the first crystal structure of a hyperthermostable ${\alpha}-CA$ from Persephonella marina EX-H1 (pmCA) in the absence and presence of competitive inhibitor, acetazolamide. The structure reveals a compactly folded pmCA homodimer in which each monomer consists of a 10-stranded ${\beta}-sheet$ in the center. The catalytic zinc ion is coordinated by three highly conserved histidine residues with an exchangeable fourth ligand (a water molecule, a bicarbonate anion, or the sulfonamide group of acetazolamide). Together with an intramolecular disulfide bond, extensive interfacial networks of hydrogen bonds, ionic and hydrophobic interactions stabilize the dimeric structure and are likely responsible for the high thermal stability. We also identified novel binding sites for calcium ions at the crystallographic interface, which serve as molecular glue linking negatively charged and otherwise repulsive surfaces. Furthermore, this large negatively charged patch appears to further increase the thermostability at alkaline pH range via favorable charge-charge interactions between pmCA and solvent molecules. These findings may assist development of novel ${\alpha}-CAs$ with improved thermal and/or alkaline stability for applications such as $CO_2$ capture and sequestration.