• Title/Summary/Keyword: i-motif

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Study on the chemical environment for conformational change of i-motif DNA by atomic force microscopy cantilever (AFM 캔틸레버를 이용한 i-motif DNA의 구조 변화에 미치는 화학적 환경에 대한 연구)

  • Jung, Hwi-Hun;Park, Jin-Sung;Yang, Jae-Moon;Lee, Sang-Woo;Eom, Kil-Ho;Kwon, Tae-Yun;Yoon, Dae-Sung
    • Journal of Sensor Science and Technology
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    • v.19 no.3
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    • pp.214-220
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    • 2010
  • Three-dimensional(3D) structure of specific DNA can be changed between two conformations under an external environmental transition such as pH and salt concentration variations. We have experimentally observed the conformational transitions of i-motif DNA using AFM cantilever bioassay. It is shown that pH change of a solvent induces the bending defleciton change of a cantilever functionalized by i-motif DNA. This indicates that cantilever bioassay enables the label-free detection of DNA structural changes upon pH change. It is implied that cantilever bioassay can be a de novo route to quantitatively understand the conformational transitions of biological molecules under environmental changes.

Cloning and Expression of Inositol Monophosphatase Gene from Streptomyces coelicolor A[3]2 (Streptomyces coelicolor A[3]2에서 Mycothiol 생합성에 관여하는 Inositol Monophosphatase 유전자의 클로닝 및 발현)

  • Kim Jin Kwon;Choi Hack Sun;Kim Seong-Jun;Kim Si Wouk
    • KSBB Journal
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    • v.19 no.6 s.89
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    • pp.462-466
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    • 2004
  • Mycothiol (MSH), a low molecular antioxidant thiol compound, was purified and analyzed from Streptomyns coelicolor A[3]2 by the monobromobimane fluorescence detection method modified by this lab. Through HPLC chromatpgram, MSH fraction was obtained following the elution time of standard MSH (donated by Dr. Robert C. Fahey). That MSH showed the highest concentration among the thiol compounds contained in the cell indicated that MSH was the key thiol compound having antioxidant activity. To understand the role of gene of inositol monophosphatase (I-1-Pase) involved in the MSH biosynthesis, it was isolated from S. coelicolor A(3)2 and cloned and overexpressed in the Escherichia coli. The expressed I-1-Pase was purified through Ni-NTA column. The soluble protein consisted of 281 amino acids, and the molecular weight was 32 kDa. I-1-Pase of S. coelicolor A(3)2 had the sequence homology with those of human and E. coli by 24 and $25\%$, respectively, and had two conserved domains (mofif A and motif B) which were typical of I-1-Pase.

Involvement of Amino Acids Flanking Glu7.32 of the Gonadotropin-releasing Hormone Receptor in the Selectivity of Antagonists

  • Wang, Chengbing;Oh, Da Young;Maiti, Kaushik;Kwon, Hyuk Bang;Cheon, Jun;Hwang, Jong-Ik;Seong, Jae Young
    • Molecules and Cells
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    • v.25 no.1
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    • pp.91-98
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    • 2008
  • The Glu/$Asp^{7.32}$ residue in extracellular loop 3 of the mammalian type-I gonadotropin-releasing hormone receptor (GnRHR) interacts with $Arg^8$ of GnRH-I, conferring preferential ligand selectivity for GnRH-I over GnRH-II. Previously, we demonstrated that the residues (Ser and Pro) flanking Glu/$Asp^{7.32}$ also play a role in the differential agonist selectivity of mammalian and non-mammalian GnRHRs. In this study, we examined the differential antagonist selectivity of wild type and mutant GnRHRs in which the Ser and Pro residues were changed. Cetrorelix, a GnRH-I antagonist, and Trptorelix-2, a GnRH-II antagonist, exhibited high selectivity for mammalian type-I and non-mammalian GnRHRs, respectively. The inhibitory activities of the antagonists were dependent on agonist concentration and subtype. Rat GnRHR in which the Ser-Glu-Pro (SEP) motif was changed to Pro-Glu-Val (PEV) or Pro-Glu-Ser (PES) had increased sensitivity to Trptorelix-2 but decreased sensitivity to Cetrorelix. Mutant bullfrog GnRHR-1 with the SEP motif had the reverse antagonist selectivity, with reduced sensitivity to Trptorelix-2 but increased sensitivity to Cetrorelix. These findings indicate that the residues flanking $Glu^{7.32}$ are important for antagonist as well as agonist selectivity.

Dynamic Profile of the Copper Chaperone CopP from Helicobacter Pylori Depending on the Bound Metals

  • Hyun, Ja-shil;Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.20 no.3
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    • pp.76-81
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    • 2016
  • Copper is an elemental ion in living organisms. CopP from Helicobacter Pylori (HpCopP) is a copper(I)-binding protein and was suggested as regulator of copper metabolism in vivo. Previously, the metal binding property of HpCopP for Ag(I), Cu(I), and Cu(II) as well as the tertiary structure of HpCopP was shown. In this study, the dynamic profiles of HpCopP depending on metal binding were studied using ${^1H}-^{15}N$ steady-state NOE analysis. The heteroNOE experiment was performed for apo-CopP or metal-bound CopP. The obtained NOE values were analyzed and compared to figure out the effect of metals on the structural flexibility of HpCopP. As a result, Ag(I) and Cu(I) ions improved the rigidity of the structure while Cu(II) ion increased the flexibility of the structure, suggesting the oxidation of the CXXC motif decreases the structural stability of HpCopP.

Repression of CCSP Expression by KLF4 (KLF4에 의한 CCSP 발현 억제)

  • Kwak, Inseok
    • Journal of Life Science
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    • v.28 no.12
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    • pp.1432-1437
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    • 2018
  • Clara cell secretory protein (CCSP) plays an important role in protecting the lungs from inflammation. This research focuses on identifying the cis-element for binding the repressor of CCSP gene expression. A DNase I footprinting experiment revealed three protected regions between -812 and -768 bp (45 bp) of the mCCSP promoter. One motif (D3: GCCTGGGAA) was 100% conserved across rat, hamster, and human. The addition of excess amounts of the D3 motif exhibited high competition within that 45 bp range in an electrophoretic mobility shift assay. However, when mutated D3 ($G{\underline{AA}}TG{\underline{TT}}AA$) was used, the competition was significantly reduced. This demonstrates that the D3 motif within that 45 bp region of the mCCSP promoter is an important site for the protein-DNA interaction. Transient transfection assays with -756 Luc resulted in highly decreased expression of CCSP than those with -812 Luc, suggesting that the 45 bp could function as a binding site for the repressor. Co-transfection of KLF4 exhibited significant repression of the -812 Luc but not the -768 Luc which clearly shows that KLF4 might function as a repressor for the CCSP gene and also suggests that the D3 motif is strongly involved in the binding of KLF4. In addition, when anti-KLF4 antibody was added, super-shifted bands were observed. This result demonstrates that KLF4 could function as a repressor by binding to this 45 bp region of the CCSP promoter and that the D3 motif might be involved in the specific binding of KLF4.

The Effect of the Oxygen Scavenging System on the pH of Buffered Sample Solutions: in the Context of Single-molecule Fluorescence Measurements

  • Kim, Sung-Eun;Lee, Il-Buem;Hong, Seok-Cheol
    • Bulletin of the Korean Chemical Society
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    • v.33 no.3
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    • pp.958-962
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    • 2012
  • In single-molecule fluorescence experiment, the oxygen scavenging system is indispensable for avoiding photo-bleaching of fluorescent dyes. Here we report that the gloxy-based oxygen scavenging system commonly used in single molecule fluorescence experiments can disturb the solution pH considerably. To track in situ pH change, we utilized the pH-sensitive conformational transition of i-motif and examined the transition with ensemble and single-molecule FRET measurements. Based on our results, we also suggested several practical remedies for the stability of the solution pH.

A Study on the Category and Classification System of Lace (레이스의 범주와 분류체계에 관한 연구)

  • Kim, Hee-Sun
    • Journal of the Korea Fashion and Costume Design Association
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    • v.16 no.4
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    • pp.117-136
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    • 2014
  • The purpose of this study is to present a classification system of the hand-made and machine-made lace according to the configuration method and re-make the category and definition of lace to consider the emergence and development of major laces techniques. The re-made category and definition of the lace is as follows. The lace usually consists of ground and motifs, however, the techniques of netting and sprang are suitable for making ground than motif, so I think it is appropriate to exclude them from the category of the lace. Many scholars are excluded openwork embroidery fabric from the category of the lace. But, an openwork embroidery fabric is the basis of a needle point lace called true lace and is consist of motif and ground. I think it is appropriate to include it in the category of the lace. I think it is also appropriate to include in the category of the lace that the eyelet embroidery fabric which mimics the openwork embroidery fabric in the machine. Lace is redefined that a fabric with openwork decoration consists of motif and ground, constructed by a variety of ways such as plaiting, twisting, looping, knotting of threads or embroidering by hand or machine. The classification of the lace is presented as follows. Hand-made lace is classified bobbin lace, needle point lace, embroidery lace, knotted lace, crochet lace, and knitting lace. Machine-made lace is classified raschel lace, leaver lace, torchon lace, and machine-made embroidery laces which include tool lace, eyelet embroidery lace, chemical lace, etc.

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NMR Studies on the Structure of Human Annexin I

  • Han, Hee-Yong;Bang, Keun-Su;Na, Doe-Sun;Lee, Bong-Jin
    • Proceedings of the Korean Biophysical Society Conference
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    • 1996.07a
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    • pp.17-17
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    • 1996
  • Annexin I is a member of the annexin family of calcium dependent phospholipid binding proteins and has anti-inflammatory activity by inhibiting phospholipase A$_2$ (PLA$_2$). Recent X-ray crystallographic study of annexin I identified six Ca$\^$2+/ binding sites, which was different types (type II, III) from the well-known EF-hand motif (type I). (omitted)

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Ecological Motif on the Salt-Water Plants of Brackish Area in Buandam (부안댐 기수역내부의 염생식물에 관한 생태적 주제성)

  • Oh Hyun-Kyung;Beon Mu-Sup;Lee Myung-Woo;Whang Bo-Chul
    • Korean Journal of Environment and Ecology
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    • v.20 no.3
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    • pp.311-318
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    • 2006
  • In an estuary, ecological dynamic is modified for a long time and memorized in soils and landscapes. As landscape memory, ecological motif is defined with dominance and rarity of vegetation. The purpose of the study is to investigate the existing vegetation character and to propose the ecological motif of this area. The present salt-water plants, community species composition and constancy degree around the estuary in Buandam watershed Buan-gun, Jeollabuk-do were analyzed. The results are as follows. The flora of the salt-water plants was listed as 16 taxa; 6 families,13 genera,14 species and 2 varieties. 5 taxa were Gramineae, 4 taxa Chenopodiaceae, and 4 taxa Comrositae. The salt-water plant communities are a total of 10 communities as listed; Zoysia sinica-Artemisia scoparia community, Phacelurus latifolius community, Artemisia scoparia Cnidium japonicum community, Limonium tetragonum -Artemisia scoparia community, Artemisia scoparia community, Suaeda japonica community, Elymus dahuricus community, Suaeda asparagoides community, Zoysia sinica community, and Zoysia sinica-Suaeda japonica community. Analyzed by the community classification species class of constancy degree, Phacelurus latifolius, Cnidium japonicum, Limonium tetragonum, Suaeda japonica, Elymus dahuricus and Suaeda asparagoides belong to I, Zoysia sinica and Artemisia scoparia to II. Elymus dahuricus, Setaria viridis var. pachystachys, Echinochloa crusgalli var. oryzicola, Phacelurus latifolius, Atriplex gmelini, Salicornia herbacea, Calystegia soldanella and Aster tripolium belong to the accompaniment species to I: Zoysia sinica, Suaeda asparagoides, Artemisia capillaris to II; Suaeda japonica, Artemisia scoparia to III ; Cnidium japonicum to IV: Limonium tetragonum, Artemisia fukudo to V. And the ecological motif is the Phacelurus latifolius community.

Detection of Lamivudine-Resistant Mutations of HBV DNA Polymerase Gene Using PCR-Direct Sequencing

  • Lee, Kyung-Ok;Lee, Hye-Jung;Byun, Ji-Young;Lee, Sung-Yeun;Kim, Jeong-Sook;Jung, Na-Young;Chung, Soo-Jin;Seong, Hye-Soon;Kim, Kyung-Tae
    • Korean Journal of Clinical Laboratory Science
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    • v.38 no.3
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    • pp.196-202
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    • 2006
  • Treatment of hepatitis B virus (HBV) with lamivudine is effective in suppressing virus replication and results in reduced inflammatory activity. However the most troublesome problem of lamivudine treatment is the emergence of lamivudine-resistant strains with amino acid substitution in the YMDD motif of DNA polymerase gene during the treatment. The aim of this study was to determine the mutation of YMDD motif (codon 552) and codon 528 in chronic HBV patients with lamivudine therapy using PCR-direct sequencing and to investigate the relationship between lamivudine mediated HBV mutation and HBeAg. HBV DNA was extracted from serum samples of HBV patients and amplified by nested PCR with two sets of primer pairs selected in HBV DNA polymerase gene. Amplified PCR product was analyzed by 2% agarose gel electrophoresis and direct sequencing. HBV mutation was detected in 124 out of 207 samples (60%). Single mutation was 50.8% for M552I, 43.5% for M552V, 5.7% for M552I/V and the L528M mutation was 67.0%. Double mutation was 43.6% for M552V/L528M, 33.1% for M552I/L528(wild type), 17.7% for M552I/L528M and 5.6% for M552I/V/L528M. Serine mutation at YMDD motif (M552S) was not found and the L528M mutation frequently accompanied M552V type. In this study, the typical difference of frequencies for HBV mutation depending on HBeAg was not found. Moreover, the PCR-direct sequencing method used in this study might be a powerful tool for the mutation study in clinical reference laboratories with high volume.

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