• 제목/요약/키워드: Anion binding

검색결과 95건 처리시간 0.022초

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

  • 백승천;김용휘;신제호;유제현
    • Journal of Dairy Science and Biotechnology
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    • 제15권1호
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    • pp.1-9
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    • 1997
  • 본 실험에서는 한우초유를 33% ammonium sulfate 포화용액으로 처리하여 조면역글로블린을 얻은 후 gel filtration, ion-exchange chromatography를 이용하여 조면역글로블린의 분리 정도를 조사하고 affinity chromatography column을 이용하여 조면역글로블린으로 부터 Ig G의 결합정도를 알아보고 Protein G Sepharose fast flow system을 이용하여 신속하게 대량으로 Ig G의 분리를 꾀하여 얻어진 Ig G를 이용하여 ELISA방법으로 항체 생성유무를 측정하였으며, 그 결과는 다음과 같다. 1. HPLC상에서 Superose 12 column을 이 용하여 한우초유의 조면역글로블린을 분리한 결과 Holstein초유의 조면역글로블린과 유사한 분리정도를 나타냈지만 약 84%의 Ig G를 한우초유의 조면역글로블린으로 부터 분리할 수 있었다. 2. Mono Q를 이용하여 HPLC에서 한우초유의 조면역글로블린을 gel filtration방법보다 짧은 시간에 분리할 수 있었지만 그다지 분리 정도가 좋지 않은 것으로 나타났다. 3. Hi-trap protein G column이 protein A sepharose CL-4B column보다 더 많은 양의 Ig G를 한우초유의 조면역글로블린으로 부터 얻을 수가 있었다. 4. Protein G Sepharose fast flow system을 이용하여 20mg의 sample양을 주입하여도 충분히 Ig G를 분리할 수 있었으며, ml 당 약 1.25mg의 Ig G를 얻을 수가 있었다. 5. ELISA방법을 이용하여 한우초유의 Ig G에 대한 항체 생성 유무를 측정한 결과, 면역화된 토끼에서 정상 토끼의 혈청에서보다 titer가 높게 나타났으므로 항체생성이 확인되었다.

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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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    • 제29권1호
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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.

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

  • 조용권
    • 생명과학회지
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    • 제18권8호
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    • pp.1036-1041
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    • 2008
  • $30{\sim}70%$ 황산암모늄 분획, Fast Q 음이온 교환수지 및 Phenyl Agarose 수지를 통해 돼지간유래의 methenyltetrahydrofolate synthetase를 정제하였으며 정제도는 119이었다. SDS-PAGE 및 HPLC를 이용한 gel permeation column chromatography에 의해 이 효소는 분자량이 23 kDa인 단량체로 확인되었다. 최적 온도와 최적 pH는 각각 $35^{\circ}C$ 와 6.5이었다. 이 효소는 tetranitromethane 및 1-ethyl-3-(3-dimethyl aminopropyl)-carbodiimide (EDC)에 의해서만 활성이 감소되었는데, 이는 티로신과 카르복실산이 효소의 활성부위에 존재한다는 것을 나타낸다. 활성부위의 pH 실험으로부터 2개의 티로신이 기질의 결합에 관여하며 하나의 카르복실기가 촉매반응에 관여한다는 사실이 관찰되었다. 따라서 이 효소는 2개의 티로신이 ATP와 5-formylTHF과 결합하며 하나의 카르복실기가 일반염기로서 촉매한다는 것을 알 수 있다.

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

  • 오길영;양수인;이완진
    • 한국대기환경학회지
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    • 제15권4호
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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
    • 한국표면공학회:학술대회논문집
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    • 한국표면공학회 2017년도 춘계학술대회 논문집
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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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미국흰불나방 지방체에서 저장단백질-1의 수용체의 특성과 부분정제 (Characterization and Partial Purification of Storage Protein-i Receptor in the Fat Body of Hyphantria cunea)

  • 채권석;여성문;김학렬
    • 한국동물학회지
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    • 제38권4호
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    • pp.490-497
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    • 1995
  • 미국흰불나방 의지방체 조직을 [35S]-메타이오닌이 포하된 배지에서 조직배양한 결과, 저장단백질-1(SP-1)의 전용기부터 용 1일 사이에 지방체로 흡수됨을 알았다. CHAPS, Triton X-100 등의 계면활성제를 농도별로 처리하여 막단백질의 용해도를 스크리닝한 뒤, anti-SP-1 polyclonal 항체를 쓴 Western blotting과 ligand blotting, 그리고 in vitro reductive methylation으로 14C을 표지한 저장단백질-1을 사용한 fluorography 등으로 1개의 수용체 밴드를 확인하였다. 1% Triton X-100으로 용해시킨 부분정제하였고, SDS-PAGE에 의해서 분자량을 측정한 결과 약 80 kDa로 나타났고 isoelectric focusing 시행 결과 등전점은 약 6.1로 계산되었다. 수용체 분자는 환원조건과 비환원조건의 차이와 전기영동 중의 온도에 따라서 SDS-PAGE상의 뚜렷한 밴드 양상의 차이를 나타내었다.

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

  • 이다혜;우희정;도정윤
    • 폴리머
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    • 제38권6호
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    • pp.801-808
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    • 2014
  • 광경화 아크릴고분자 필름에서 이온이동에 관한 연구를 하였다. 고분자에 결합된 아민기와 카르복실산 작용기의 함량을 조절하여 수소이온과 수산화이온의 이동에 적합한 필름을 제조하였다. 수소이온 이동 검출을 위해 p-methylred(PMR) 유도체를 사용하였고 수산화이온 검출을 위해 페놀프탈레인 유도체를 합성하여 각각 고분자에 도입하였다. 수소이온의 이동은 아민기의 함량이 많을수록 빠르게 진행되었고 수산화이온은 카르복실산의 함량이 높을수록 빠르게 이동하였다. 수소이온 이동은 필름표면과 내부로 빠르게 진행하였고 PMR 포함 필름의 흡수스펙트럼을 통해 관찰되었다. 산 용액에 감응하는 필름을 사용하여 색상변화의 가역성을 관찰하였고 연속 50회를 실시하는 동안 정확히 재현되었다. 구리(II) 이온은 카르복실산 작용기의 함량이 높은 필름에서 빠르게 이동하였다. 로다민이 도입된 필름에서 구리이온의 배위로 로다민의 고리열림 반응이 진행되었고 광흡수 및 발광특성 측정을 통해 구리이온의 이동을 추적하였다.

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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    • 제19권9호
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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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    • 제31권12호
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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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    • 제42권6호
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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.