• Title/Summary/Keyword: Dichroism

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Structural stability of CD1 domain of human mitotic checkpoint serine/threonine-protein kinase, Bub1

  • Kim, Hyun-Hwi;Song, Hyun-Kyu;Lee, Bong-Jin;Park, Sung Jean
    • Journal of the Korean Magnetic Resonance Society
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    • v.19 no.2
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    • pp.88-94
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    • 2015
  • Bub1 is one of the spindle checkpoint proteins and plays a role in recruitment of the related proteins to kinetochore. Here, we studied the structural characteristic of the evolutionarily conserved 160 amino acid region in the N-terminus (hBub1 CD1), using Circular Dichroism (CD) and NMR. Our CD results showed that hBub1 CD1 is a highly helical protein and its structure was affected by pH: as pH was elevated to basic pH, the helical propensity increased. This could be related to the surface charge of the hBub1 CD1. However, the structural change did not largely depend on the salt concentration, though the thermal stability a little increased. The previous NMR analysis revealed that the hBub1 CD1 adopts eight helices, which is consistent with the CD result. Our result would be helpful for evaluating the molecular mechanism of the hBub1 CD1 and protein-protein interactions.

Catalytic Properties of Monomeric Species of Brain Pyridoxine-5'-phosphate Oxidase

  • Kwon, Oh-Shin;Choi, Soo-Young
    • BMB Reports
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    • v.34 no.1
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    • pp.21-27
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    • 2001
  • The structural stability of brain pyrydoxine-5'-phosphate (PNP) oxidase and the catalytic properties of the monomeric species were investigated. The unfolding of brain pyridoxine-5'-phosphate (PNP) oxidase by guanidine hydrochloride (GuHCl) was monitored by means of fluorescence and circular dichroism spectroscopy Reversible dissociation of the dimeric enzyme into subunits was attained by the addition of 2 M GuHCl. The perturbation of the secondary structure under the denaturation condition resulted in the release of the cofactor FMN. Separation of the processes of refolding and reassociation of the monomeric species was achieved by the immobilization method. Dimeric PNP oxidase was immobilized by the covalent attachment to Affi-gel 15 without any significant lass of its catalytic activity. Matrix-bound monomeric species were obtained from the reversible refolding processes. The matrix bound-monomer was found to be catalytically active, possessing only a slightly decreased specific activity when compared to the refolded dimeric enzyme. In addition, limited chymotrypsin digestion of the oxidase yields two fragments of 12 and 161 kDa with a concomitant increase of catalytic activity The catalytically active fragment was isolated by ion exchange chromatography and analyzed for association of two subunits using the FPLC gel filtration analysis. The retention time indicated that the catalytic fragment of 16 kDa behaves as a compact monomer. Taken together, these results are consistent with the hypothesis that the native quaternary structure of PNP oxidase is not a prerequisite for catalytic function, but it could play a role in the regulation.

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Influence of the Hydrophobic Amino Acids in the N- and C-Terminal Regions of Pleurocidin on Antifungal Activity

  • Lee, June-Young;Lee, Dong-Gun
    • Journal of Microbiology and Biotechnology
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    • v.20 no.8
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    • pp.1192-1195
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    • 2010
  • To investigate the influence of the N- or C-terminal regions of pleurocidin (Ple) peptide on antifungal activity, four analogs partially truncated in the N- or C-terminal regions were designed and synthesized. Circular dichroism (CD) spectroscopy demonstrated that all the analogs maintained an alpha-helical structure. The antifungal susceptibility testing also showed that the analogs exhibited antifungal activities against human fungal pathogens, without hemolytic effects against human erythrocytes. The result further indicated that the analogs had discrepant antifungal activities [Ple>Ple (1-22)>Ple (4-25)>Ple (1- 19)>Ple (7-25)] and that N-terminal deletion affected the activities much more than C-terminal deletion. Hydrophobicity [Ple>Ple (1-22)>Ple (4-25)>Ple (1-19)> Ple (7-25)] was thought to have been one of the consistent factors that influenced these activity patterns, rather than the other primary factors like the helicity [Ple>Ple (4-25) >Ple (1-22)>Ple (1-19)>Ple (7-25)] or the net charge [Ple=Ple (4-25)=Ple (7-25)>Ple (1-22)=Ple (1-19)] of the peptides. In conclusion, the hydrophobic amino acids in the N-terminal region of Ple is more crucial for antifungal activity than those in the C-terminal region.

Protective Effect of Biological Osmolytes against Heat- and Chaotropic Agent-Induced Denaturation of Bacillus licheniformis γ-Glutamyl Transpeptidase

  • Lo, Huei-Fen;Chi, Meng-Chun;Lin, Min-Guan;Lan, Yuan-Gin;Wang, Tzu-Fan;Lin, Long-Liu
    • Journal of Microbiology and Biotechnology
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    • v.28 no.9
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    • pp.1457-1466
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    • 2018
  • In the present study, the stabilizing effect of four different biological osmolytes on Bacillus licheniformis ${\gamma}$-glutamyl transpeptidase (BlGGT) was investigated. BlGGT appeared to be stable under temperatures below $40^{\circ}C$, but the enzyme retained less than 10% of its activity at $60^{\circ}C$. The tested osmolytes exhibited different degrees of effectiveness against temperature inactivation of BlGGT, and sucrose was found to be the most effective among these. The use of circular dichroism spectroscopy for studying the secondary structure of BlGGT revealed that the temperature-induced conformational change of the protein molecule could be prevented by the osmolytes. Consistently, the molecular structure of the enzyme was essentially conserved by the osmolytes at elevated temperatures as monitored by fluorescence spectroscopy. Sucrose was further observed to counteract guanidine hydrochloride (GdnHCl)-and urea-induced denaturation of BlGGT. Taken together, we observed evidently that some well-known biological osmolytes, especially sucrose, make a dominant contribution to the structural stabilization of BlGTT.

Pharmaceutical Studies on the Inclusion Complexes of Non-Steroidal Antiinflammatory Drugs with ${\beta}-Cyclodextrin$ (I) (비(非)Steroid 성소염약물(性消炎藥物)과 ${\beta}-Cyclodextrin$과의 Inclusion Complex에 관(關)한 약제학적(藥劑學的) 연구(硏究) (제1보)(第一報))

  • Han, Kun;Lee, Min-Hwa;Kim, Shin-Keun
    • Journal of Pharmaceutical Investigation
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    • v.13 no.1
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    • pp.10-22
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    • 1983
  • The interactions of ${\alpha}-cyclodextrin({\alpha}-CyD)$ and ${\beta}-cyclodextrin({\beta}-CyD)$ with several non-steroidal antiinflammatory drugs were studied on the effects of ${\alpha}-$ and ${\beta}-CyD$ on the solubility of the drugs in aqueous medium. Indoprofen, niflumic acid, alclofenac, and naproxen were chosen as representatives of antiinflammatory drugs. The solubility of all drugs studied increased with the addition of ${\beta}-CyD$, while not with glucose or ${\alpha}-CyD$. The increase of the solubility with ${\beta}-CyD$ was considered due mainly to the formation of inclusion complexes between ${\beta}-CyD$ and drugs. From the solubility data, the apparent stability constants K of the complex could be calculated. Ultraviolet absorption and circular dichroism confirmed the inclusion of indoprofen, niflumic acid and naproxen with ${\beta}-CyD$ in the molar ratio of 1 : 1. Inclusion complexes in solid powder form were obtained by the freeze-drying method and the inclusion formation was confirmed again by infrared, diffential thermal analysis, and X-ray diffraction measurements.

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Structural Characterization of Growth-Related Translationally Controlled Tumor Protein P23

  • Lee, Bong-Jin;Hong, Yoon-Hun;Park, Sang-Ho;Lee, Kyunglim
    • Journal of the Korean Magnetic Resonance Society
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    • v.5 no.1
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    • pp.46-55
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    • 2001
  • P23, a translationally controlled turner protein is involved in the interleukin-4 secretion from human basophils and is also known to be an IgE-dependent histamine-releasing factor. However, the precise physiological function and structure of P23 have not been elucidated. In the current study, we constructed the optimal expression and purification protocol of P23 and investigated the secondary structure and structural stability in various conditions. Circular dichroism (CD) investigation showed that the secondary structure of P23 adopts mainly a P-sheet conformation. CD spectroscopy and differential scanning calorimetry revealed that P23 is fairly stable in the pH range of neutral and mild-basic conditions and in the temperature range of 10 - 50$\^{C}$. Since the thermal stability and the P-sheet content of P23 were decreased by the addition of Ca$\^$2+/ ion, it could be suggested that Ca$\^$2+/ion induces structural change by partially destabilizing the structure of P23. In addition various H experiments were monitored to solve the aggregation of P23. Den results will provide the preliminary structural information about P23.

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A Study on the Attractive Items of Hanok in Urban Area focused on Preceding Studies (선행연구에 나타난 도시한옥의 매력 요소에 관한 연구)

  • Min, Sae-Rom;Kim, Tai-Young
    • Journal of the Korean Institute of Rural Architecture
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    • v.14 no.3
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    • pp.61-68
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    • 2012
  • The purpose of this paper is to derive attractive items of 'Hanok' in order to preserve 'Hanok' in urban area, focused on the 13 preceding studies which have got in respect of image, satisfaction, preference, awareness and advantage of 'Hanok' in urban area revolved around the latest studies. This study is analysis of questionnaire items and results of 13 preceding studies. And attractive items are classified with general and overall survey items, advantage of 'Hanok' in urban area, and image of Hanok-density urban area. These can also be classified with 5 items which are space, health, material, form, and image of 'Hanok' in urban area. As a result, Space items are composed with 'use of a space', 'cosiness', 'garden' and 'floor and ondol(korean floor heating system). Health items are composed with 'lighting and ventialtion', 'a sense of the season' and 'natural material'. Material items are composed with 'natural beauty', 'wooden', 'changhoji(traditional Korean paper made from mulberry bark for doors and windows) and hanji(traditional Korean paper handmade from mulberry trees), 'rafter' and 'new materials'. Form items are composed with 'attractive appearance', 'simply decoration' and 'traditional elements'. Image items are composed with 'crowding', 'traditionality and historicity', 'dichroism' and 'warmth'.

The Effect of Cr Dosage on FePt Nanoparticle Formation

  • Won, C.;Keavney, D.J.;Divan, R.;Bader, S.D.
    • Journal of Magnetics
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    • v.11 no.4
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    • pp.182-188
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    • 2006
  • The search for high-density recording materials has been one of most active and vigorous field in the field of magnetism. $FePt-L1_{0}$ nanoparticle has emerged as a potential candidate because of its high anisotropy. In this paper, we provide an overview of recent work at Argonne National Laboratory that contributes to the ongoing dialogue concerning the relation between structure and properties of the FePt nanoparticle system. In particular we discuss the ability to control structure and properties via dosing with Cr. Cr-dosed FePt films were grown via molecular beam epitaxy and annealed at $550^{\circ}C$ in an ultrahigh vacuum chamber, and were studied with the surface magneto-optic Kerr effect (SMOKE), scanning electron microscopy (SEM) and x-ray magnetic circular dichroism (XMCD). We found that small dosage of Cr helps to generate $L1_{0}$ phase FePt magnetic nanoparticles with small size, defined shape and regular spatial distribution on MgO (001) substrate. The nanostructures are ferromagnetic with high magnetic coercivity (${\sim}0.9T$) and magnetic easy axis in the desired out-of-plane orientation. We also show that controlling the lateral region where nanostructures exist is possible via artificial patterning with Cr.

Structural Effects of the GXXXG Motif on the Oligomer Formation of Transmembrane Domain of Syndecan-4

  • Song, Jooyoung;Kim, Ji-Sun;Choi, Sung-Sub;Kim, Yongae
    • Bulletin of the Korean Chemical Society
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    • v.34 no.12
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    • pp.3577-3585
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    • 2013
  • Syndecan-4 (heparan sulfate proteoglycan), biologically important in cell-to-cell interactions and tumor suppression, was studied through mutation of the GXXXG motif of its transmembrane domain (Syd4-TM), a motif which governs dimerization. The expression and purification of the mutant (mSyd4-TM) were optimized here to assess the function of the GXXXG motif in the dimerization of Syd4-TM. mSyd4-TM was obtained in M9 minimal media and its oligomerization was identified by SDS PAGE, Circular Dichroism (CD) spectroscopy, mass spectrometry and NMR spectroscopy. The mutant, unlike Syd4-TM, did not form dimers and was observed as monomers. The GXXXG motif of Syd-4TM was shown to be an important structural determinant of its dimerization.

Cell Selectivity of an Antimicrobial Peptide Melittin Diastereomer with D-amino Acid in the Leucine Zipper Sequence

  • Zhu, Wan Long;Nan, Yong Hai;Hahm, Kyung-Soo;Shin, Song-Yub
    • BMB Reports
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    • v.40 no.6
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    • pp.1090-1094
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    • 2007
  • Melittin (ME), a linear 26-residue non-cell-selective antimicrobial peptide, displays strong lytic activity against bacterial and human red blood cells. To design ME analogue with improved cell selectivity, we synthesized a melittin diastereomer (ME-D) with D-amino acid in the leucine zipper sequence (Leu-6, Lue-13 and Ile-20). Compared to ME, ME-D exhibited the same or 2-fold higher antibacterial activity but 8-fold less hemolytic activity. Circular dichroism analysis revealed that ME-D has much less $\alpha$-helical content in $\alpha$-helical content in the presence of zwitterionic EYPC/cholesterol (10 : 1, w/w) liposomes compared to negatively charged EYPE/EYPG (7 : 3, w/w) liposomes. The blue shift of the fluorescence emission maximum of ME-D in zwitterionic EYPC/cholesterol (10 : 1, w/w) liposomes was much smaller than in negatively charged EYPE/EYPG (7 : 3, w/w) liposomes. These results suggested that the improvement in therapeutic index/cell selectivity of ME-D is correlated with its less permeability to zwitterionic membranes.