• Title/Summary/Keyword: In-folding Structure

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Unfolding of Ervatamin C in the Presence of Organic Solvents: Sequential Transitions of the Protein in the O-state

  • Sundd, Monica;Kundu, Suman;Dubey, Vikash Kumar;Jagannadham, Medicherla V.
    • BMB Reports
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    • v.37 no.5
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    • pp.586-596
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    • 2004
  • The folding of ervatamin C was investigated in the presence of various fluorinated and non-fluorinated organic solvents. The differences in the unfolding of the protein in the presence of various organic solvents and the stabilities of O-states were interpreted. At pH 2.0, non-fluorinated alkyl alcohols induced a switch from the native $\alpha$-helix to a $\beta$-sheet, contrary to the $\beta$-sheet to $\alpha$-helix conversion observed for many proteins. The magnitude of ellipticity at 215 nm, used as a measure of $\beta$-content, was found to be dependent on the concentration of the alcohol. Under similar conditions of pH, fluorinated alcohol enhanced the intrinsic a-helicity of the protein molecule, whereas the addition of acetonitrile reduced the helical content. Ervatamin C exhibited high stability towards GuHCl induced unfolding in different O-states. Whereas the thermal unfolding of O-states was non-cooperative, contrary to the cooperativity seen in the absence of the organic solvents under similar conditions. Moreover, the differential scanning calorimetry endotherms of the protein acquired at pH 2.0 were deconvoluted into two distinct peaks, suggesting two cooperative transitions. With increase in pH, the shape of the thermogram changed markedly to exhibit a major and a minor transition. The appearance of two distinct peaks in the DSC together with the non-cooperative thermal transition of the protein in O-states indicates that the molecular structure of ervatamin C consists of two domains with different stabilities.

The Physical Characteristics of Chinese Hand-made and Machine-made Noodles (시중 중화면의 수타면과 기계면의 물리적 특성)

  • Kim, Sung-Su;Yoon, Jang-Ho;Lee, Seung-Ju
    • Journal of the East Asian Society of Dietary Life
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    • v.18 no.1
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    • pp.80-86
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    • 2008
  • Both hand-made and machine-made Chinese noodles are popular in Korea. In this study, each type was evaluated in terms of its physical properties to rigorously determine for consumers which one has better qualities. The noodles were instrumentally measured for color, size, moisture content, density, viscoelasticity, and cutting force. The behaviors of the noodles were visually observed during cooking, and sensory evaluations were performed with the cooked noodles. The hand-made raw noodles were less dense, had higher moisture content, and generated more bubbles during cooking than the machine-made noodles. This indicated that the hand-made noodles contained more entrapped air, thereby resulting in the above physical characteristics. The change in noodle size after cooking was greater in the hand-made noodles, indicating that more entrapped air in expansion escaped during cooking and was replaced by water. The cutting force and viscoelasticity of the hand-made noodles were lower, and were controlled by viscous properties, respectively. These results agreed with the fact that the hand-made noodles had higher moisture content and lower density. In the sensory evaluation, the hand-made noodles presented lower hardness, but higher elasticity. It was inferred that the hand-made noodle dough underwent repeated processes of folding and extending, resulting in better developed of the gluten structure. Consequently, the hand-made noodles were determined to be different than the machine-made noodles in terms of instrumental measurements and sensory observations, suggesting that the hand-made noodles had superior textural properties.

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Biophysical Studies Reveal Key Interactions between Papiliocin-Derived PapN and Lipopolysaccharide in Gram-Negative Bacteria

  • Durai, Prasannavenkatesh;Lee, Yeongjoon;Kim, Jieun;Jeon, Dasom;Kim, Yangmee
    • Journal of Microbiology and Biotechnology
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    • v.28 no.5
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    • pp.671-678
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    • 2018
  • Papiliocin, isolated from the swallowtail butterfly (Papilio xuthus), is an antimicrobial peptide with high selectivity against gram-negative bacteria. We previously showed that the N-terminal helix of papiliocin (PapN) plays a key role in the antibacterial and anti-inflammatory activity of papiliocin. In this study, we measured the selectivity of PapN against multidrug-resistant gram-negative bacteria, as well as its anti-inflammatory activity. Interactions between Trp2 of PapN and lipopolysaccharide (LPS), which is a major component of the outer membrane of gram-negative bacteria, were studied using the Trp fluorescence blue shift and quenching in LPS micelles. Furthermore, using circular dichroism, we investigated the interactions between PapN and LPS, showing that LPS plays critical roles in peptide folding. Our results demonstrated that Trp2 in PapN was buried deep in the negatively charged LPS, and Trp2 induced the ${\alpha}$-helical structure of PapN. Importantly, docking studies determined that predominant electrostatic interactions of positively charged arginine residues in PapN with phosphate head groups of LPS were key factors for binding. Similarly, hydrophobic interactions by aromatic residues of PapN with fatty acid chains in LPS were also significant for binding. These results may facilitate the development of peptide antibiotics with anti-inflammatory activity.

A Case Study on the Polar Low Developed over the Sea Near Busan on 11~12 February 2011 (2011년 2월 11~12일 부산 근해에서 발달한 극저기압에 대한 사례연구)

  • Lee, Jae Gyoo;Kim, Hae-Min;Kim, Yu-Jin
    • Atmosphere
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    • v.26 no.2
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    • pp.301-319
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    • 2016
  • The evolutionary process of the polar low, which caused the heavy snowfall in the East Coast area on 11~12 February 2011, was investigated to describe in detail using synoptic weather charts, satellite imageries, and ERA (European Centre for Medium-Range Weather Forecasts Re-Analysis) -Interim reanalysis data. It was revealed that 1) the polar low was generated over the sea near Busan where a large cyclonic shear in the inverted trough branched from the parent low existed, 2) during the developing and mature stages, there was a convectively unstable region in the lower layer around the polar low and its south side, 3) the polar low was developed in the region where the static stability in the 500~850 hPa layer was the lowest, 4) the result from the budget analysis of the vorticity equation indicated that the increase in the vorticity at the lower atmosphere, where the polar low was located, was dominated mainly by the stretching term, 5) the warm core structure of the polar low was identified in the surface-700 hPa layer during the mature stage, 6) there was a close inverse relationship between a development of the polar low and the height of the dynamic tropopause over the polar low, and 7) for generation and development of the polar low, large-scale circulation systems, such as upper cold low and its combined short wave trough, major low (parent low), and polar air outbreak, should be presented, indicating that the polar low has the nature of the baroclinic disturbance.

Development of a Novel Subunit Vaccine Targeting Fusobacterium nucleatum FomA Porin Based on In Silico Analysis

  • Jeong, Kwangjoon;Sao, Puth;Park, Mi-Jin;Lee, Hansol;Kim, Shi Ho;Rhee, Joon Haeng;Lee, Shee Eun
    • International Journal of Oral Biology
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    • v.42 no.2
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    • pp.63-70
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    • 2017
  • Selecting an appropriate antigen with optimal immunogenicity and physicochemical properties is a pivotal factor to develop a protein based subunit vaccine. Despite rapid progress in modern molecular cloning and recombinant protein technology, there remains a huge challenge for purifying and using protein antigens rich in hydrophobic domains, such as membrane associated proteins. To overcome current limitations using hydrophobic proteins as vaccine antigens, we adopted in silico analyses which included bioinformatic prediction and sequence-based protein 3D structure modeling, to develop a novel periodontitis subunit vaccine against the outer membrane protein FomA of Fusobacterium nucleatum. To generate an optimal antigen candidate, we predicted hydrophilicity and B cell epitope parameter by querying to web-based databases, and designed a truncated FomA (tFomA) candidate with better solubility and preserved B cell epitopes. The truncated recombinant protein was engineered to expose epitopes on the surface through simulating amino acid sequence-based 3D folding in aqueous environment. The recombinant tFomA was further expressed and purified, and its immunological properties were evaluated. In the mice intranasal vaccination study, tFomA significantly induced antigen-specific IgG and sIgA responses in both systemic and oral-mucosal compartments, respectively. Our results testify that intelligent in silico designing of antigens provide amenable vaccine epitopes from hard-to-manufacture hydrophobic domain rich microbial antigens.

Effects of Boshimgeonbi-tang on Gene Expression in Hypothalamus of Immobilization-stressed Mouse (보심건비탕(補心健脾湯) 투여가 Stress 유발 Mouse의 Hypothalamus 유전자 발현에 미치는 영향)

  • Lee Seoung-Hee;Chang Gyu-Tae;Kim Jang-Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.19 no.6
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    • pp.1585-1593
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    • 2005
  • The genetic effects of restraint stress challenge on HPA axis and the therapeutic effect of Boshimgeonbi-tang on the stress were studied with cDNA microarray analyses, RT-PCR on hypothalamus using an immobilization-stress mice as an animal model. Male CD-1 mice were restrained in a tightly fitted and ventilated vinyl holder for 2hrs once a day, and this challenge was repeated for seven· consecutive days. In the change of body weight it showed that the Boshimgeonbi-tang is effected recovery on weight loss caused by the immobilization-stress. Seven days later, total RNA was extracted from the organs of the mouse, body-labeled with $CyDye^{TM}$ fluorescence dyes and then hybridized to CDNA microarray chip. Scanning and analyzing the array slides were carried out using GenePix4000 series scanner and GenePix $Pro^{TM}$ analyzing program, respectively. The expression profiles of 109 genes out of 6000 genes on the chip were significantly modulated in hypothalamus by the immobilization stress. Energy metabolism-, lipid metabolism-, apoptosis-, stress protein, transcriptional factor, and signal transduction-related genes were transcriptionally activated whereas DNA repair-, protein biosysthesis-, and structure integrity-related genes were down-regulated in hypothalamus. The 58 genes were up-regulated by the mRNA expression folds of 1.5 to 7.9. and the 51 genes were down-regulated by 1.5 - 5.5 fold. The 11 genes among them were selected to confirm the expression profiles by RT-PCR. The mRNA expression levels of Tnfrsf1a (apoptosis), Calm2 (cell cycle), Bag3 (apoptosis), Ogg1 (DNA repair), Aatk (apoptosis), Dffa (apoptosis), Fkbp5 (protein folding) were restored to the normal one by the treatment of Boshimgeonbi-tang.

Analysis of the Rrigidity and the Vibration of Flat Corrugated Plates (주름판의 강성해석 및 진동해석)

  • Han, B.K.;Chung, K.;Yoo, S.Y.
    • Transactions of the Korean Society of Mechanical Engineers
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    • v.15 no.1
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    • pp.11-18
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    • 1991
  • Stiffened plate structure, which is generally used in the various structural design to develope the load carrying capacity, is classified in two groups; one is the plate stiffened with stiffeners, the other is corrugated plate. In the studies on those structures, the studies on the stiffened plates with stiffeners have been much studied with both quantities and qualities according to requirements of the minimum-weight structural design and the development in many industrial fields, especially automobile, ship and aerospace fields, but the studies on the corrugated plates are undeveloped in comparison with the stiffened plates, and also the analytical stiffness on the corrugated plates remains as the imperfect. In the present studies, the analytical method on the stiffness of corrugated plates made by folding is proposed, and the stiffness equation of corrugated plates with some angle is derived and generalized. The purpose of the present study is to contribute to the design of corrugated plates and to determine the optimum aspect ratio for parameters that decide the aspect of corrugated plates.

A Study on Characteristics of Jointed Rock Masses and Thermo-hydro-mechanical Behavior of Rock Mass under High Temperature (방사성 폐기물 저장을 위한 불연속 암반의 특성 및 고온하에서의 암반의 수리열역학적 상호작용에 관한 연구)

  • 이희근;김영근;이희석
    • Tunnel and Underground Space
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    • v.8 no.3
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    • pp.184-193
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    • 1998
  • In order to dispose radioactive wastes safely, it is needed to understand the mechanical, thermal, fluid behavior of rockmass and physico-chemical interactions between rockmass and water. Also, the knowledge about mechanical and hydraulic properties of rocks is required to predict and to model many conditions of geological structure, underground in-situ stress, folding, hot water interaction, intrusion of magma, plate tectonics etc. This study is based on researches about rock mechanics issues associated with a waste disposal in deep rockmass. This paper includes the mechanical and hydraulic behavior of rocks in varying temperature conditions, thermo-hydro-mechanical coupling analysis in rock mass and deformation behavior of discontinuous rocks. The mechanical properties were measured with Interaken rock mechanics testing systems and hydraulic properties were measured with transient pulse permeability measuring systems. In all results, rock properties were sensitive to temperature variation.

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cDNA Cloning, Sequence Analysis and Molecular Modeling of a New Peptide from the Scorpion Buthotus saulcyi Venom

  • Nikkhah, Maryam;Naderi-Manesh, Hossein;Taghdir, Majid;Talebzadeh, Mehdi;Sadeghi-Zadeh, Majid;Schaller, Janatan;Sarbolouki, Mohamad N.
    • BMB Reports
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    • v.39 no.3
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    • pp.284-291
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    • 2006
  • In this study, the cDNA of a new peptide from the venom of the scorpion, Buthotus saulcyi, was cloned and sequenced. It codes for a 64 residues peptide (Bsaul1) which shares high sequence similarity with depressant insect toxins of scorpions. The differences between them mainly appear in the loop1 which connects the $\beta$-strand1 to the $\alpha$-helix and seems to be functionally important in long chain scorpion neurotoxins. This loop is three amino acids longer in Bsaul1 compared to other depressant toxins. A comparative amino acid sequence analysis done on Bsaul1 and some of $\alpha$-, $\beta$-, excitatory and depressant toxins of scorpions showed that Bsaul1 contains all the residues which are highly conserved among long chain scorpion neurotoxins. Structural model of Bsaul1 was generated using Ts1 (a $\beta$-toxin that competes with the depressant insect toxins for binding to $Na^+$ channels) as template. According to the molecular model of Bsaul1, the folding of the polypeptide chain is being composed of an anti-parallel three-stranded $\beta$-sheet and a stretch of $\alpha$-helix, tightly bound by a set of four disulfide bridges. A striking similarity in the spatial arrangement of some critical residues was shown by superposition of the backbone conformation of Bsaul1 and Ts1.

Kinetic Studies of Peptidylprolyl cis-trans Isomerase from Porcine Spleen

  • Kim, Soo-Ja;Lee, Chan
    • BMB Reports
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    • v.29 no.6
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    • pp.519-524
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    • 1996
  • Peptidylprolyl cis-trans isomerase (PPlase) catalyzes the cis-trans isomerization of prolyl peptide and facilitates the folding of cellular proteins and peptides. PPlase consists of two distinct immunophilins, each specifically binding to the immunosupressive drug cyclosporin A (CsA) or FK506, respectively. A PPlase was isolated and partially purified from porcine spleen. The molecular weight of porcine spleen PPlase was determined to be ~14,000 on the basis of SDS-PAGE. The purified enzyme was strongly inhibited by FK506, but not by CsA. The inhibition constant and the true concentration of enzyme preparations were determined by active site titration using the tight binding inhibitor FK506: $K_{i}=18.7$ nM and $E_{t}=172$ nM. The equilibrium ratio of conformer. [cis]/[trans], of prolyl peptide substrates (N-Suc-Ala-Xaa-Pro-Phe-p-NA) in anhydrous trifluoroethanol/LiCl solvent system varied from 0.24 to 0.85 depending on the nature of Xaa. Overall. in this solvent-salt system, the populations of the cis conformer of substrates in equilibrium are higher than in an aqueous solution so that the substantial error caused by high background absorption can be reduced. The reactivities of porcine spleen PPlase are shown to be highly sensitive to changes in the structure of substrates. Thus, $k_{cat}/K_m$ value for the most reactive substrate (Xaa Leu) is $4.007+10^{6}M^{1}s^{1}$ and, is 2,636 fold higher than that for the least reactive peptide substrate tested, Xaa=Glu.

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