• Title/Summary/Keyword: Single-molecule

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The Crystal Structure of an Iondine Sorption Complex of Dehydrated Calcium and Silver Exchanged Zeolite A ($Ag^+$이온과 $Ca^{2+}$이온으로 치환한 제올라이트 A를 탈수한 후 요오드를 흡착한 결정구조)

  • Bae, Myung-Nam;Kim, Yang;Kim, Un-Sik
    • Korean Journal of Crystallography
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    • v.6 no.2
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    • pp.118-124
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    • 1995
  • The crystal structure of an iodine sorption complex of vacumm-dehydrated Ag+ and Ca2+ exchanged zeolite A(a=12.174(3)Å) has been determined at 21℃ by single-crystal X-ray diffraction techniques in the cubic space group Pm3m. The crystal was prepared by flow method for three days using exchange solution in solution in which mole ratio of AgNO3 and Ca(NO3)2 was 1:150 with total concentration of 0.05 M. The complex was prepared by dehydration at 360℃ and 2×10-6 Torr for 2 days, followed by exposure to about 14.3 Torr of iodine vaporat 80℃ for 24 hours. Full-matrix least-squares refinement converged to the final error indices of R1=0.082, R2=0.068 using 122 reflections for which I > 3σ(I). Two Ag+ ions, 1.1 Ag+ ions, and 4.45 Ca2+ ions per unit cell are located on three different three-fold axes associated with 6-ring oxygens. Two Ag+ ions per unit cell are in the large cavity, 1.399(4)Å from the (111) plane of three oxygens. Another 1.1 Ag+ ions are found at opposite sites. Six iodine molecules are sorbed per unit cell. Each I2 molecule approaches a framework oxide ion axially (O-I=3.43(2)Å, I-I=2.92Å, I-I-O;166.1(3)°), by a charge transfer complex interaction. Two Ag+ ions make a close approach to the iodine molecules (Ag-I ; 2.73(2)Å).

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The Solvent-Independent Structure of 6-(2-pyridyl)-3, 5-hexadiyn-1-ol (6-(2-pyridyl)-3, 5-hexadiyn-1-ol의 용매 비의존 분자구조)

  • 서일환;이진호
    • Korean Journal of Crystallography
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    • v.6 no.1
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    • pp.36-42
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    • 1995
  • Two types of single crystals of the title compound [6-(2-pyridyl)-3, 5-hexadiyn-ol, PyHxD] were obtained by solution of n-hexane/CH2C12 and n-hexane/Et2O, and their molecular conformations are proved identical in spite of different of space groups; C22H18N2O2 (I), Mr=343.70, Monoclinic, Pa, a=14.595(2), b=5.413(2), c=12.218(2)Å, β=96.86(1)°, V=958.3Å3, Z=2, Dx=1.19 Mgm-3, λ(MoKα)=0.71069Å, μ=0.072mm-1, F(000)=360.0, T=292K, R=0.104 for 756 unique observed reflections. An asymmetric unit contains a dimer connected by two N-H…O intermolecular hydrogen bonds. C11H9NO (II), Mr=171.85, Monoclinic, P21/a, a=14.611(2), b=5.423(6), c=12.191(2)Å, β=96.89(1)°, V=959.0Å3, Z=4, Dx=1.19 Mgm-3, λ(MoKα)=0.71069Å, μ=0.072mm-1, F(000)=360.0, T=293K, R=0.066 for 824 unique observed reflection. The structural asymmetric unit contains a molecule, but two N-H…O hydrogen bonds related by controsymmetry make the molecules form a dimer. In both structure, the dihedral angle between the planar pyridyl ring and the plane defined by C(10)-C(11)-O connected by linear diyne chain is approximately normal, and the molecules are stacked along b-axis with the unit repeat of b-axis.

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Solid-phase PEGylation for Site-Specific Modification of Recombinant Interferon ${\alpha}$-2a : Process Performance, Characterization, and In-vitro Bioactivity (재조합 인터페론 알파-2a의 부위 특이적 수식을 위한 고체상 PEGylation : 공정 성능, 특성화 및 생물학적 활성)

  • Lee, Byung-Kook;Kwon, Jin-Sook;Lee, E.K.
    • KSBB Journal
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    • v.21 no.2
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    • pp.133-139
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    • 2006
  • In 'solid-phase' PEGylation, the conjugation reaction occurs as the proteins are attached to a solid matrix, and thus it can have distinct advantages over the conventional, solution-phase process. We report a case study: rhIFN-${\alpha}$-2a was first adsorbed to cation exchange resin and then N-terminally PEGylated by aldehyde mPEG of 5, 10, and 20 kD through reductive alkylation. After the PEGylation, salt gradient elution efficiently recovered the mono-PEGylate in a purified form from the unwanted species such as unmodified IFN, unreacted PEG, and others. The mono-PEGylation and its purification were integrated in a single chromatographic step. Depending on the molecular weight of the mPEG aldehyde used, the mono-PEGylation yield ranged 50-64%. We could overcome the major problems of random, or uncontrollable, multi-PEGylation and the post-PEGylation purification difficulties associated with the solution-phase process. N-terminal sequencing and MALDI-TOF MS confirmed that a PEG molecule was conjugated only to the N-terminus. Compared with the unmodified IFN, the mono-PEGylate showed the reduced anti-viral activity as measured by the cell proliferation assay. The bioactivity was reduced more as the higher molecular weight PEG was conjugated. Immunoreactivity, evaluated indirectly by antibody binding activity using a surface plasmon resonance biosensor, also decreased. Nevertheless, trypsin resistance as well as thermal stability was considerably improved.

Bacterial Toxin-antitoxin Systems and Their Biotechnological Applications (박테리아의 toxin-antitoxin system과 생명공학기술 응용)

  • Kim, Yoonji;Hwang, Jihwan
    • Journal of Life Science
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    • v.26 no.2
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    • pp.265-274
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    • 2016
  • Toxin-antitoxin (TA) systems are ubiquitous genetic modules that are evolutionally conserved in bacteria and archaea. TA systems composed of an intracellular toxin and its antidote (antitoxin) are currently classified into five types. Commonly, activation of toxins under stress conditions inhibits diverse cellular processes and consequently induces cell death or reversible growth inhibition. These effects of toxins play various physiological roles in such as regulation of gene expression, growth control (stress response), programmed cell arrest, persister cells, programmed cell death, phage protection, stabilization of mobile genetic elements or postsegregational killing of plasmid-free cells. Accordingly, bacterial TA systems are commonly considered as stress-responsive genetic modules. However, molecule screening for activation of toxin in TA system is available as development of antimicrobial agents. In addition, cytotoxic effect induced by toxin is used as effective cloning method with antitoxic effect of antitoxin; consequently cells containing cloning vector inserted a target gene can survive and false-positive transformants are removed. Also, TA system is applicable to efficient single protein production in biotechnology industry because toxins that are site-specific ribonuclease inhibit protein synthesis except for target protein. Furthermore, some TA systems that induce apoptosis in eukaryotic cells such as cancer cells or virus-infected cells would have a wide range of applications in eukaryotes, and it will lead to new ways of treating human disease. In this review, we summarize the current knowledge on bacterial TA systems and their applications.

Exocyclic GpC DNA methyltransferase from Celeribacter marinus IMCC12053 (Celeribacter marinus IMCC12053의 외향고리 GpC DNA 메틸트랜스퍼라아제)

  • Kim, Junghee;Oh, Hyun-Myung
    • Korean Journal of Microbiology
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    • v.55 no.2
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    • pp.103-111
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    • 2019
  • DNA methylation is involved in diverse processes in bacteria, including maintenance of genome integrity and regulation of gene expression. CcrM, the DNA methyltransferase conserved in Alphaproteobacterial species, carries out $N^6$-adenine or $N^4$-cytosine methyltransferase activities using S-adenosyl methionine as a co-substrate. Celeribacter marinus IMCC12053 from the Alphaproteobacterial group was isolated from a marine environment. Single molecule real-time sequencing method (SMRT) was used to detect the methylation patterns of C. marinus IMCC12053. Gibbs motif sampler program was used to observe the conversion of adenosine of 5'-GANTC-3' to $N^6$-methyladenosine and conversion of $N^4$-cytosine of 5'-GpC-3' to $N^4$-methylcytosine. Exocyclic DNA methyltransferase from the genome of strain IMCC12053 was chosen using phylogenetic analysis and $N^4$-cytosine methyltransferase was cloned. IPTG inducer was used to confirm the methylation activity of DNA methylase, and cloned into a pQE30 vector using dam-/dcm- E. coli as the expression host. The genomic DNA and the plasmid carrying methylase-encoding sequences were extracted and cleaved with restriction enzymes that were sensitive to methylation, to confirm the methylation activity. These methylases protected the restriction enzyme site once IPTG-induced methylases methylated the chromosome and plasmid, harboring the DNA methylase. In this study, cloned exocyclic DNA methylases were investigated for potential use as a novel type of GpC methylase for molecular biology and epigenetics.

Hematopoietic Stem Cells and Bone Marrow Microenvironment: Current and Emerging Concepts (골수 미세환경에서 조혈줄기세포의 기능조절에 대한 고찰- 현재 및 새로운 개념)

  • Lee, Won Jong;Park, Seong Hyun;Park, Jun Hee;Oh, Seong Hwan;Lee, Dongjun
    • Journal of Life Science
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    • v.32 no.6
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    • pp.468-475
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    • 2022
  • The functional distinction between stem and progenitor cells is well established in several tissues, particularly in the blood. There, hematopoietic stem cells preserve their self-renewal potential and reconstitution ability in the bone marrow niche. Bone marrow represents a unique setting in which to examine how stroma influences tissue function. It was the setting in which the experimental definition of a niche was first provided in mammalian stem cell biology and where clear evidence for non-cell-autonomous oncogenesis was first defined. The relationship between bone and blood is ancient as all animals since the divergence of fish that have bones and blood, make blood in their bones. This long coevolution engendered complex interrelationships, including the first proposed and first experimentally defined niche for stem cells in mammals. Multiple bone marrow stromal cell types serve as regulators of hematopoiesis, and the dysfunction of some causes myelodysplasia and leukemia. However, no comprehensive atlas of stromal subpopulations exists. Therefore, we think these data point to something of importance, such as how the needs and challenges of the organism become translated down to distinct cell types that critically govern specific functions within tissues and do so at the level of a single molecule. We think this will be of broad interest to those focusing on systems biology and the physiology of organisms, particularly those seeking a molecular basis for understanding cell and tissue behavior. We summarized the current and emerging concepts of hematopoietic stem cells and bone marrow niche.

Structural Study of Selenium Sorption Complex of Fully Dehydrated, Partially Ca2+-exchanged Zeolite A (완전히 탈수되고 부분적으로 칼슘 이온으로 교환된 제올라이트 A의 셀레늄 수착 화합물의 구조 연구)

  • Kim, Hu Sik;Park, Jong Sam;Lim, Woo Taik
    • Korean Journal of Mineralogy and Petrology
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    • v.33 no.3
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    • pp.251-258
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    • 2020
  • Single crystal of fully dehydrated and partially Ca2+-exchanged zeolites A (|Ca4Na4|[Si12Al12O48]-LTA) was brought into contact with Se in fine pyrex capillary at 523 K for 5 days. Crystal structure of Se-sorbed |Ca4Na4|[Si12Al12O48]-LTA has been determined by single-crystal X-ray diffraction techniques at 294 K in the cubic space group $Pm{\bar{3}}m$ (a = 12.2787(13) Å). The crystal structure of yellow |Ca4Na4Se4|[Si12Al12O48]-LTA has been refined to the final error indices of R1/wR2 = 0.0960/0.3483 with 327 reflections for which Fo > 4s(Fo). In this structure, 4 Na+ and 4 Ca2+ ions fill every 6-ring site: These ions are all found at three crystallographic positions, on 3-fold axes equipoints of opposite 6-rings. Selenium atoms are found at three crystallographically distinct positions: 2 Se atoms per unit cell at Se(1) are located opposite 6-rings in the sodalite cavity (Se(1)-Na(1) = 2.53(5) Å) and 1 at Se(2) opposite 4-rings (Se(2)-O(1) = 2.76(10) Å) and 1 at Se(3) opposite 6-rings in the large cavity (Se(3)-Na(1) = 2.48(5) Å). Two molecular of Se2 (Se(1)-Se(1) = 2.37(7) or 2.90(8) Å and Se(2)-Se(3) = 2.91(5) ) Å) are found in all sodalite cavity and large cavity. Other clusters such as Se4 and Se8 could be existed in large cavity. The inter-selenium distances turned out to be longer that of gases Se2 molecule.

Effect of Antibody Immobilization Method to Magnetic Micro Beads on its Immunobinding Characteristics (자성 미세입자에의 항체 고정화 방법이 면역결합반응에 미치는 영향)

  • Choi, Hyo Jin;Hwang, Sang Youn;Jang, Dae Ho;Cho, Hyung Min;Kang, Jung Hye;Seong, Gi Hun;Choo, Jae Bum;Lee, Eun Kyu
    • Korean Chemical Engineering Research
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    • v.44 no.1
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    • pp.65-72
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    • 2006
  • Recent technical advances in the biorecognition engineering and the microparticle fabrication may enable us to develop the single step purification using magnetic particle, because of its simplicity, efficacy, ease of automation, and process economics. In this study, we used commercial magnetic particles from Seradyn, Inc. (Indianapolis, USA). It was ca. 2.8 micron in diameter, consisted of polystyrene core and magnetite coating, and its surface had carboxyl groups. The model, capture protein was IgG and anti-IgG was used as the ligand molecule. We studied the different surfaces ('nude', ester-activated, and anti-IgG coated) for their biorecognition of IgG. At a high pH condition, we could reduce non-specific binding. Also anti-IgG immobilized magnetic particle could capture IgG more selectively. We attempted 'oriented immobilization' of anti-IgG, in which the polysaccharides moiety near the C-terminus was selectively oxidized and linked to the hydrazine-coated MP, to improve the efficacy of biorecognitive binding. Using this method, the IgG capturing ability was improved by ca. 2 fold. From the binary mixture of the IgG-insulin, IgG could be more selectively captured. In summary, the oriented immobilization of oxidized anti-IgG proved to be as effective as the streptavidin-biotin system and yet simpler and cost-effective. This immobilization method can find its applications in protein biochips and biotargeting.

The Value of ICAM-1 Expression and the Soluble ICAM-1(sICAM-1) Level as a Marker of Activity in Sarcoidosis: The Relationship Between the ICAM-1 Level and the Clinical Course of the Disease (유육종증의 활동성 지표로서의 ICAM-1)

  • Kim, Dong-Soon;Paik, Sang-Hoon;Shim, Tae-Sun;Lim, Chae-Man;Lee, Sang-Do;Koh, Youn-Suck;Kim, Woo-Sung;Kim, Won-Dong
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.1
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    • pp.116-127
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    • 1998
  • Background: The natural course of sarcoidosis is variable from spontaneous remission to significant morbidity or death. So the assessment of disease activity is important but no single parameter was generally accepted as a good marker. Recently several studies suggested that adhesion molecules, especially ICAM-1 can be a marker, but there are some controversies. And only few data are available about the relationship of ICAM-1 with clinical follow-up course. Methods: We measured the expression of adhesion molecules on BAL cells by flow cytometry and the level of soluble ICAM-1(sICAM-1) in serum and BALF at the time of diagnosis in 12 patients with active disease and 7 inactive sarcoidosis(5 male, 14 female, mean age: $39.4{\pm}10.7$ years, mean follow-up : $20{\pm}15$ months). Follow-up clinical course were compared with the changes in serum sICAMA-1 level and the adhesion molecule on BAL cells. Results: In the patients with active disease, the ICAM-1 on AM(RMFI: $3.68{\pm}1.71$) and sICAM-1 level in serum($582{\pm}193$ng/ml) and BAL fluid($47.8{\pm}16.5$ng/ml) were all higher than those of 7 inactive disease(RMFI: $1.89{\pm}0.75$, p=0.0298, serum: $294{\pm}117$ ng/ml, p=0.0049, BALF: $20.9{\pm}8.3$ ng/ml). In the active sarcoidosis, ICAM-1 on AM(RMFI : $1.51{\pm}0.84$) and serum sICAM-1 were decreased after the therapy($250{\pm}147$ ng/ml) but no significant change was noted in inactive disease. Also we found the initial ICAM-1 on AM and serum sICAM-1 had a significant correlation with the degree of improvement in PFT after the therapy. During the follow-up, the disease relapsed in 4 patients after the discontinuation of steroid and the serum sICAM-1 level went-up again at the time of relapse. Conclusion: Our data suggest that the serum sICAM-1 level and the ICAM-1 expression on AM can be a good marker of disease activity and also a predictor of outcome in sarcoidosis.

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