• 제목/요약/키워드: structural conformation

검색결과 141건 처리시간 0.024초

NMR and Circular Dichroism Studies on Human CD99 Transmembrane Domain

  • Kim, Hai-Young;Shin, Joon;Shin, Young-Kee;Park, Seong-Hoe;Lee, Weon-Tae
    • 한국자기공명학회논문지
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    • 제7권1호
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    • pp.37-45
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    • 2003
  • Human CD99 is a ubiquitous 32-kDa transmembrane protein encoded by mic2 gene. Recently it has been reported that expression of a splice variant of CD99 transmembrane protein (Type I and Type II) increases invasive ability of human breast cancer cells. To understand structural basis for cellular functions of CD99 Type II, we have initiated studies on hCD99$\^$TMcytoI/ using circular dichroism (CD) and multi-dimensional NMR spectroscopy. CD spectrum of hCD99$\^$TMytoI/ in the presence of 200mM DPC and CHAPS displayed an existence ${\alpha}$-helical conformation, showing that it could form an ${\alpha}$-helix under membrane environments. In addition, we have found that the cytoplasmic domain of CD99 would form symmetric dimmer in the presence of transmembrane domain. Although it has been rarely figured out the correlation between structure and functional mechanism of hCD99$\^$TMcytoI/, the dimerization or oligomerization would play an important role in its biological function.

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NMR Structural Study of the 3'-T.G Mismatched DNA Decamer Duplex Containing the T-T (6-4) Adduct

  • Lee, Joon-Hwa;Park, Yun-Jeong;Park, Byong-Seok
    • 한국자기공명학회논문지
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    • 제3권1호
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    • pp.60-70
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    • 1999
  • The pyrimidine(6-4) pyrimidone photoproduct [(6-4) adduct] is one of the major photoproducts induced by UV irradiation of DNA and occurs at TpT sites. The (6-4) adduct is highly mutagenic and specific during translesion replication. The marked preference for insertion of A opposite the 5'-T and G opposite the 3--T of the (6-4) adducts leads to a predominantly 3'-T\longrightarrowC transition with 85% replicating error rate. In order to obtain insight into the origin of 3'-T\longrightarrowC transition induced by the (6-4) adduct, we have performed one - and two-dimensional NMR experiment. The 3'-Tof the (6-4) lesion forms the stable hydrogen bonding to the imino proton of an opposed G, which stabilizes the overall helix and diminishes the highly distorted conformation caused by the (6-4) lesion in the (6-4)/AA duplex. We proposed that the greater insertion of a G over an A opposite the 3'-T of the (6-4) lesion These results may account for the greater preference for the insertion of a G over a A opposite the 3'-T of the (6-4) lesion. Thus this insertion leads to the highly specific 3'-T\longrightarrowC multation at the (6-4) lesion site.

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NMR에 의한 입체구조에 관한 연구 (제1보). ${\alpha},{\beta}$-Unsaturated Phenylhydrazones의 Conformation과 Configuration에 대하여 (Structural Studies by NMR (Ⅰ). Conformations and Configurations of ${\alpha},{\beta}$-Unsaturated Phenylhydrazones)

  • 이학기;임용진;최정도
    • 대한화학회지
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    • 제18권3호
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    • pp.215-220
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    • 1974
  • 저자들은 tetrahedrane생성의 전구체인 t-butenedial bistosylhydrazone의 입체적인 구조를 연구할 목적으로 이의 유사한 구조인 ${\alpha},{\beta}$위치에 이중결합을 가지고 있는 몇 가지 phenylhydrazone을 합성하고 이들의 형태 및 배위를 nmr spectra의 해석으로부터 구명했다. 벤젠용매와 aliphatic solvent(CC14, DMSO)에서의 chemical shift의 차이로 부터 이들 물질이 syn-configuration을 가지고 있음을 알았으며 spin-spin coupling constant 로부터 이들 물질의 형태를 고찰하였다.

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CNT의 도입에 의한 β-phase PVDF의 형성과 CNT/PVDF 복합막에서의 압전성능 개선 (Formation of β-phase PVDF by Introduction of CNTs in the CNT/PVDF Composite Film and Resulting Improvement of Piezoelectric Performance)

  • 임영택;이선우
    • 한국전기전자재료학회논문지
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    • 제29권11호
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    • pp.712-715
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    • 2016
  • In this paper, we fabricated flexible CNT/PVDF piezoelectric composite device by introducing CNTs (carbon nanotubes) into PVDF (poly-vinylidene fluoride) solution using spray coating technique. Flexible PEDOT:PSS conducting polymer was used as electrodes. We tried to improve the piezoelectric performance from the CNT/PVDF composite film by increasing the portion of the ${\beta}$-phase PVDF in the film. We confirmed the structural conformation of the CNT/PVDF composite film as a function of CNT concentration by using FT-IR (fourier transform infra-red). As increasing CNT concentration, portion of the ${\beta}$-phase PVDF and resulting piezoelectric performance increased in the CNT/PVDF composite film. We found that CNTs introduced were played as seeds for formation of the ${\beta}$-phase PVDF in the CNT/PVDF composite film and resulting improvement of the piezoelectric performance.

TECHNIQUES FOR INTERGRANULAR CRACK FORMATION AND ASSESSMENT IN ALLOY 600 BASE AND ALLOY 182 WELD METALS

  • LEE, TAE HYUN;HWANG, IL SOON;KIM, HONG DEOK;KIM, JI HYUN
    • Nuclear Engineering and Technology
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    • 제47권1호
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    • pp.102-114
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    • 2015
  • Background: A technique developed to produce artificial intergranular stress corrosion cracks in structural components was applied to thick, forged alloy 600 base and alloy 182 weld metals for use in the qualification of nondestructive examination techniques for welded components in nuclear power plants. Methods: An externally controlled procedure was demonstrated to produce intergranular stress corrosion cracks that are comparable to service-induced cracks in both the base and weld metals. During the process of crack generation, an online direct current potential drop method using array probes was used to measure and monitor the sizes and shapes of the cracks. Results: A microstructural characterization of the produced cracks revealed realistic conformation of the crack faces unlike those in machined notches produced by an electrodischarge machine or simple fatigue loading using a universal testing machine. Conclusion: A comparison with a destructive metallographic examination showed that the characteristics, orientations, and sizes of the intergranular cracks produced in this study are highly reproducible.

CRYSTAL STRUCTURE OF tRNA ($m^1$ G37) METHYLTRANSFERASE

  • Ahn, Hyung-Jun;Lee, Byung-Ill;Yoon, Hye-Jin;Yang, Jin-Kuk;Suh, Se-Won
    • 한국결정학회:학술대회논문집
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    • 한국결정학회 2003년도 춘계학술연구발표회
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    • pp.17-17
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    • 2003
  • tRNA (m¹ G37) methyltransferase (TrmD) catalyze s the trans for of a methyl group from S-adenosyl-L-methionine (AdoMet) to G/sup 37/ within a subset of bacterial tRNA species, which have a residue G at 36th position. The modified guanosine is adjacent to and 3' of the anticodon and is essential for the maintenance of the correct reading frame during translation. We have determined the first crystal structure of TrmD from Haemophilus influenzae, as a binary complex with either AdoMet or S-adenosyl-L-homocysteine (AdoHcy), as a ternary complex with AdoHcy/phosphate, and as an apo form. The structure indicates that TrmD functions as a dimer (Figure 1). It also suggests the binding mode of G/sup 36/G/sup 37/ in the active site of TrmD and catalytic mechanism. The N-terminal domain has a trefoil knot, in which AdoMet or AdoHcy is bound in a novel, bent conformation. The C-terminal domain shows a structural similarity to DNA binding domain of trp or tot repressor. We propose a plausible model for the TrmD₂-tRNA₂ complex, which provides insights into recognition of the general tRNA structure by TrmD (Figure 2).

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The active site and substrate binding mode of 1-aminocyclopropane-1- carboxylate oxidase of Fuji apple (Malus domesticus L.) determined by site directed mutagenesis and comparative modeling studies

  • Ahrim Yoo;Seo, Young-Sam;Sung, Soon-Kee;Yang, Dae-Ryook;Kim, Woo-Tae-K;Lee, Weontae
    • 한국생물물리학회:학술대회논문집
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    • 한국생물물리학회 2003년도 정기총회 및 학술발표회
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    • pp.70-70
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    • 2003
  • Active sites and substrate bindings of 1-aminoxyclopropane-1-carboxylate oxidase (MD-ACO1) catalyzing the oxidative conversion of ACC to ethylene have been determined based on site-directed mutagenesis and comparative modeling methods. Molecular modeling based on the crystal structure of Isopenicillin N synthase (IPNS) provided MD-ACO1 structure. MD-ACO1 protein folds into a compact jelly roll shape, consisting of 9 ${\alpha}$-helices, 10 ${\beta}$-strands and several long loops. The MD-ACO1/ACC/Fe(II)/Ascorbate complex conformation was determined from automated docking program, AUTODOCK. The MD-ACO1/Fell complex model was consistent with well known binding motif information (HIS177-ASP179-HIS234). The cosubstrate, ascorbate is placed between iron binding pocket and Arg244 of MD-ACO1 enzyme, supporting the critical role of Arg244 for generating reaction product. These findings are strongly supported by previous biochemical data as well as site-directed mutagenesis data. The structure of enzyme/substrate suggests the structural mechanism for the biochemical role as well as substrate specificity of MD-ACO1 enzyme.

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Comparison of Surface and Core Peptide Fraction from Apo B-100 of Human LDL (Low Density Lipoprotein)

  • Cho, Hyun-Mi;Shin, Seung-Uon;Kim, Tae-Woong
    • Preventive Nutrition and Food Science
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    • 제4권2호
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    • pp.145-151
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    • 1999
  • Apolipoprotein B-100 (apo B-100) is an important component in plasma low density lipoproteins (LDL). It function as the ligand for the LDL receptor in peripheral cells. The LDLs are removed from the circulation by both high-affinity receptor-mediated and receptor-independant pathways. LDLs are heterogeneous in their lipid content, size and density and certain LDL subspecies increase risk of atherosclerosis due to differences in the conformation of apo B in the particle. In the present study , surface and core peptide fraction of Apo B-100 have been characterized by comparing peptide-mapping and fluorescence spectroscopy. Surface fragments of apo B-100 were generated by digestion of LDL with either trypsin , pronase, or pancreatin elastase. Surface fractions were fractionated on a Sephadex G-50 column. The remaining core fragments were delipidated and redigested with the above enzymes, and the resulting core peptides were compared with surface peptides. Results from peptide-mapping by HPLC showed pronase-digestion was more extensive than trypsin -digestion to remove surface peptide fraction from LDL. Fluorescence spectra showed that core fractions contained higher amount of tryptophan than surface fractions, and it indicated that core fraction wa smore hydrophobic than surface fractions. A comparison of the behavior of the core and surface provided informations about the regions of apo B-100 involved in LDL metabolism and also about the structural features concerning the formation of atherosclerosis.

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Isolation of Proteins that Specifically Interact with the ATPase Domain of Mammalian ER Chaperone, BiP

  • Chung, Kyung-Tae;Lee, Tae-Ho;Kang, Gyong-Suk
    • Biotechnology and Bioprocess Engineering:BBE
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    • 제8권3호
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    • pp.192-198
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    • 2003
  • BiP, immunoglobulin binding protein, is an ER homologue of Hsp70. However, unlit other Hsp70 proteins, regulatory protein(s) for BiP has not been identified. Here, we demo strafed the presence of potential regulatory proteins for BiP using a pull -down assay. Since BiP can bind any unfolded protein, only the ATPase domain of BiP was used for the pull -down assay in order to minimize nonspecific binding. The ATPase domain was cloned to produce recombinant protein, which was then conjugated to CNBr-activated agarose. The structural conformation and ATP hydrolysis activity of the recombinant ATPase domain were similar to those of the native protein, light proteins from metabolically labeled mouse plasmacytoma cells specifically bound to the recombinant ATPase protein. The binding of these proteins was inhibited by excess amounts of free ATPase protein, and was dependent on the presence of ATP. These proteins were eluted by ADP. Of these proteins, Grp170 and BiP where identified. while the other were not identified as known ER proteins, from Western blot analyses. The presence of the ATPase-binding proteins for BiP was first demonstrated in this study, and our data suggest similar regulatory machinery for BiP may exist in the ER, as found in prokaryotes and other cellular compartments.

Site-directed Mutagenesis of Arginine 13 Residue in Human Glutathione S-Transferase P1-1

  • Koh, Jong-Uk;Cho, Hyun-Young;Kong, Kwang-Hoon
    • Bulletin of the Korean Chemical Society
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    • 제28권5호
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    • pp.772-776
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    • 2007
  • In order to study the role of residue in the active site of glutathione S-transferase (GST), Arg13 residue in human GST P1-1 was replaced with alanine, lysine and leucine by site-directed mutagenesis to obtain mutants R13A, R13K and R13L. These three mutant enzymes were expressed in Escherichia coli and purified to electrophoretic homogeneity by affinity chromatography on immobilized GSH. Mutation of Arg13 into Ala caused a substantial reduction of the specific activity by 10-fold. Km GSH, Km DCNB and Km EPNP values of R13A were approximately 2-3 fold larger than those of the wild type. Mutation of Arg13 into Ala also significantly affected I50 values of S-methyl-GSH that compete with GSH and ethacrynic acid, an electrophilic substrate-like compound. These results appeared that the substitution of Arg13 with Ala resulted in significant structural change of the active site. Mutation of Arg13 into Leu reduced the catalytic activity by approximately 2-fold, whereas substitution by Lys scarcely affected the activity, indicating the significance of a positively charged residue at position 13. Therefore, arginine 13 participates in catalytic activity as mainly involved in the construction of the proper electrostatic field and conformation of the active site in human GST P1-1.