• Title/Summary/Keyword: Binding property

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Structural Studies on RUNX of Caenorhabditis elegans by Spectroscopic Methods

  • Son, Woo-Sung;Kim, Jong-Wan;Ahn, Hee-Chul;Park, Sung-Jean;Bae, Suk-Chul;Lee, Bong-Jin
    • Journal of the Korean Magnetic Resonance Society
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    • v.6 no.1
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    • pp.54-68
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    • 2002
  • PEBP2/CBF (Polyomavirus Enhancer-core Binding Protein 2/Core Binding Factor), represents a new family of heterodimeric transcription factor. Those members play important roles in hematopoiesis and osteogenesis in mouse and human. PEBP2/CBF is a sequence-specific DNA binding protein. Each member of the PEBP2/CBF family of transcription factors is composed of two subunits, ${\alpha}$ and ${\beta}$. The evolutionarily conserved 128 amino acid region in ${\alpha}$ subunit has been called the Runt domain, which harbors two different activities, the ability to bind DNA and interact with the ${\beta}$ subunit. Recently, cDNA clones encoding the C. elegans Runt domain were isolated by screening a cDNA library. This gene was referred to run (Runt homologous gene). In this study, the basic experiments for the structural characterization of RUN protein were performed using spectroscopic methods. We have identified the structural properties of RUN using bioinformatics, CD and NMR. The limit temperature of the structural stability was up to 60$^{\circ}C$ with irreversible thermal process, and the structure of RUN seems to adopt ${\alpha}$ helices and one or more ${\beta}$ sheet or turn. The degree of NMR peak dispersion and intensity was increased by addition of glycine. Therefore, glycine could be used to alleviate the aggregation property of RUN in NMR experiment.

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Oligomeric Structures Determine the Biochemical Characteristics of Human Nucleoside Diphosphate Kinases

  • Kim, Sun-Young;Song, Eun-Joo;Chang, Keun-Hye;Kim, Eun-Hee;Chae, Suhn-Kee;Lee, Han-Soo;Lee, Kong-Joo
    • BMB Reports
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    • v.34 no.4
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    • pp.355-364
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    • 2001
  • Major human Nucleoside diphosphate kinases (NDPKs) exist as hetero-oligomers, consisting of NDPK-A and NDPK-B, rather than homo-oligomer. To investigate their biological function depending on the oligomeric structure in vivo, we characterized the biochemical properties of cellular NDPK. Cellular NDPKs, which are made up of a unique combination of isoforms, were purified from human erythrocyte and placenta. We found that cellular NDPK and recombinant isoforms NDPKs have their own distinct biochemical properties in autophosphorylation, stability toward heat or urea, and DNA binding. Cellular NDPK was found to have unique characteristics rather than the expected additive properties of recombinant isoforms. The mutations in the dimeric interface of NDPK-B (R34G, N69H or K135L) caused defective DNA binding and simultaneously reduced the enzymatic stability These results suggest that the oligomeric interaction could play a major role in the stability of catalytic domain and might be related to the regulation of various cellular functions of NDPK.

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Expression, Purification and Functional and structural relationship of pyruvate dehydrogenase phosphatase

  • Kim, Young-Mi;Jung, Ki-Hwa
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 2002.07a
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    • pp.236-236
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    • 2002
  • Pyruvate dehydrogenase phosphatase (PDP) is a mitochondrial protein serine/threonine phosphatase that catalyzes the dephosphorylation and concomitant reactivation of the pyruvate dehydrogenase componant of the pyruvate dehydrogenase complex (PDC). PDP consists of a Mg$\^$+2/ -dependent and Ca$\^$+2)-stimulated catalytic subunit (PDPc) of Mr 52,600 and a FAD-containing regulatory subunit (PDPr) of Mr 95.600. Catalytic subunit of pyruvate dehydrogenase phosphatase (PDPc) has been suggested to have three major functional domains such as dihydrolipoamide acetyltransferase(E$_2$)-binding domain, regulatory subunit of PDP(PDPr)-binding domain, and calcium-binding domain. In order to identify functional domains, recombinant catalytic subunit of pyruvate dehydrogenase phosphatase (rPDPc) was expressed in E. coli JM101 and purified to near homogeneity using the unique property of PDPc: PDPm binds to the inner lipoyl domain (L$_2$) of E$_2$ of pyruvate dehydrogenase complex (PDC) in the presence of Ca$\^$+2/, not under EGTA. PDPc was limited-proteolysed by trypsin, chymotrypsin, Arg-C, and elastase at pH7.0 and 30$^{\circ}C$ and N-terminal analysis of the fragment was done. Chymotrypsin, trypsin, and elastase made two major framents: N-terminal large fragment, approx. 50kD and C-terminal small fragment, approx. 0 kDa. Arg-C made three major fragments: N-terminal fragment, approx. 35 kD, and central fragment, approx. 15 kD, and C-terminal fragment, approx. 10 kD. This study strongly suggest that PDPc consists of three major functional domains. However, further study should be necessary to identify the functional role.

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Relative potency of antihistaminics for $H_1$-and muscarinic receptors (항 히스타민제의 $H_1$ 수용체와 무스카린 수용체에 대한 상대적 역가)

  • 이신웅;박영주;이정수
    • YAKHAK HOEJI
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    • v.37 no.4
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    • pp.397-407
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    • 1993
  • The muscarinic antagonist l-[benzilic-4,4'-$^3H$]quinuclidinyl benzilate([$^3H$]QNB) bound to a single class of muscarinic receptor with high affinity in guinea pig ileal membranes. The $K_{D}$ and B$_{max}$ values for [$^3H$]QNB calculated from analysis of saturation isotherms were 54 pM and 156fmol/mg, respectively. H$_{1}$-blockers inhibited [$^3H$]QNB binding to ileal membranes with $K_{i}$ values ranged from 0.008 $\mu{M}$ to 1.6 $\mu{M}$. The pseudo-Hill coefficients of H$_{1}$-blockers for inhibition of [$^3H$]QNB binding to the ileal membranes were close to unit. The $K_{i}$ values for H$_{1}$-blockers were similar to the $K_{M}$ values calculated by Schild plot of functional data obtained from inhibition of the carbachol-induced contraction in guinea-pig ileum, suggesting that binding of H$_{1}$-blockers vs [$^3H$]QNB in ileal membranes represents an interaction with a receptor of physiological relevance. The $K_{H}$ values of H$_{1}$-blockers for H$_{1}$-receptor estimated from inhibition of the histamine-induced contraction were the range of 0.15 nM to 56.5 nM. The $K_{M}$/K$_{H}$ ratio of H$_{1}$-blockers varied over a wide range of 3 to 2300. Thus, the antihistaminic potencies of H$_{1}$-blockers do not correlate with their antimuscarinic potencies, which suggest that antihistamines have different antimuscarinic potencies in therapeutic blood levels causing similar antiallergic effect. Among 13 traditional antihistaminics examined in this study, drug having the highest and the lowest $K_{M}$/K$_{H}$ ratio is triprolidine and diphenidol, respectively. The present results demonstrate that the antimuscarinic property of antihistamines is not necessary for their antiallergic effect, and data on the affinity of antihistamines for muscarinic and H$_{1}$-receptors can be an important parameter in the selection and evaluation of these drugs.

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Physicochemical Properties of Potato Starches Hydroxypropylated with Propylene Oxide (Propylene Oxide로 하이드록시프로필화시킨 감자 전분의 이화학적 특성)

  • 배수경;이선자;김미라
    • Journal of the East Asian Society of Dietary Life
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    • v.7 no.4
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    • pp.519-526
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    • 1997
  • Physicochemical properties of potato starches with degree of substitution of 0-0.2645 hydroxypropylated with propylene oxide were examined. Hydorxypropyl group content of the starch was proportional to propylene oxide concentration. Water binding capacity increased but pH decreased with the increase of degree of substitution. Blue value and iodine absorption property decreased with the increase of hydroxypropylation. Differential scanning calorimeter showed that hydroxypropylation lowered the gelatinization temperature of the starch. Relative crystallinity in X-ray diffraction patterns decreased and starch granule observed by scanning electron microscopy was destroyed by hydroxypropylation. This result implied that high level hydroxypropylation affected the crystal region as well as amorphous region of starch.

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Improvement on the Functional Properties of Gelatin Prepared from the Yellowfin Sole Skin by Precipitation with Ethanol (알코올처리에 의한 각시가자미껍질 젤라틴의 기능성 개선)

  • Kim, Jin-Soo;Lee, Eung-Ho
    • Korean Journal of Food Science and Technology
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    • v.26 no.6
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    • pp.683-689
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    • 1994
  • With a view to utilizing effectively fish skin wastes from marine manufactory, a gelatin solution extracted from yellowfin sole skin was fractionated by precipitation with ethanol, and then the functional and physico-chemical properties for the fractionated gelatin were determined. Ethanol was added up to 50% of ethanol content to a gelatin solution extracted from yellowfin sole skin, then the mixture was left to stand at $0^{\circ}C$ for 12 hours. Finally, the precipitates were dried by hot-air ($40^{\circ}C$). The gel strength and melting point of a 10% gel of gelatin prepared from yellowfin sole skin by precipitation with ethanol has 322.4g and $23.3^{\circ}C$, respectively. The physico-chemical properties of the ethanol treated fish skin gelatin were superior to those of fish skin gelatin prepared without ethanol treatment. Besides, the functional properties of the ethanol treated gelatin were lower in solubility and higher in water holding capacity, oil binding capacity, emulsifying activity, emulsifying stability, foam expansion and foam stability than those of pork skin gelatin sold on market as well as gelatin prepared without ethanol treatment. It may be concluded, from these results, that the fish skin gelatin prepared by precipitation with ethanol can be effectively utilized as a human food by improving the functional properties.

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Fabrication and Electrical Characteristics of Ferredoxin Self-Assembled Layer for Biomolecular Electronic Device Application

  • NAM YUN SUK;CHOI JEONG-WOO
    • Journal of Microbiology and Biotechnology
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    • v.16 no.1
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    • pp.15-19
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    • 2006
  • A ferredoxin adsorbed hetero self-assembled layer was fabricated on chemically modified Au substrate, 4-Aminothiophenol (4-ATP) was deposited onto Au substrate and then N-succinimidyl-3-[2-pyridyldithio] propionate (SPDP) was adsorbed on the 4-ATP layer, since SPDP was used as a bridging molecule for ferredoxin adsorption, Ferredoxin/SPDP/4-ATP structured hetero layer was constructed because of strong chemical binding of ferredoxin, SPDP, and 4-ATP, The surface of the ferredoxin-adsorbed SPDP/4-ATP layer was observed by scanning tunneling microscopy, The hetero film formation was verified by surface plasmon resonance measurement. The current flow and rectifying property based on the scanning tunneling spectroscopy I-V characteristics was achieved in the proposed hetero layer. Thus, the hetero layer structure of ferredoxin functioned as a molecular diode with rectifying property, The proposed molecular diode can be usefully applied for the development of molecular scale electronic devices.

Functional and Film-forming Properties of Fractionated Barley Proteins

  • Cho, Seung-Yong;Rhee, Chul
    • Food Science and Biotechnology
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    • v.18 no.4
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    • pp.889-894
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    • 2009
  • Barley proteins are expected to have unique functional properties due to their high content of alcohol soluble protein, hordein. Since the barley proteins obtained by conventional isoelectric precipitation method cannot represent hordein fraction, barley proteins were fractionated to albumin, globulin, glutelin, and hordein with respect to extraction solvents. Functional properties and film-forming properties of solubility-fractionated barley proteins were investigated to explore their potential for human food ingredient and industrial usage. The 100 g of total barley protein comprised 5 g albumin, 23 g globulin, 45 g glutelin, and 27 g hordein. Water-binding capacities of barley protein isolates ranged from 140-183 mL water/100 g solid. Hordein showed the highest oil absorption capacity (136 mL oil/100 g), and glutelin showed the highest gelation property among the fractionated proteins. In general, the barley protein fractions formed brittle and weak films as indicated by low tensile strength (TS) and percent elongation at break (E) values. The salt-soluble globulin fraction produced film with the lowest TS value. Although films made from glutelin and hordein were dark-colored and had lower E values, they could be used as excellent barriers against water transmission.

Microbial Transglutaminase Improves the Property of Meat Protein and Sausage Texture Manufactured with Low-quality Pork Loins

  • Katayama, K.;Chin, K.B.;Yoshihara, S.;Muguruma, M.
    • Asian-Australasian Journal of Animal Sciences
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    • v.19 no.1
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    • pp.102-108
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    • 2006
  • Microbial transglutaminase (MTGase) was investigated to determine whether it was an effective binding agent for the processing of low-quality pork loins. MTGase especially promoted the coagulation of myosin heavy chain (MHC). However, the effect of MTGase on MHC from low-quality meat was less than that from the normal meat when the reaction time was not enough. The breaking strength of the heat-induced gel made of myosin B from low-quality meat with MTGase was lower than that of normal meat. Sausage made with low-quality meat with MTGase did not exhibit improved hardness, as compared to that made with normal meat. Results of this study indicated that use of low-quality meat in the manufacture of sausage was feasible to get textural property equal to that of normal meat sausage, when a half or more of the raw material was normal meat and MTGase was used in the sausage.

Current Issues & Prospects of International Space Law

  • Zwaan, Tanja Masson
    • The Korean Journal of Air & Space Law and Policy
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    • v.25 no.1
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    • pp.237-259
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    • 2010
  • This paper first gives a brief overview of the history of space law making in the international geopolitical context and recalls some of the main principles as elaborated in the framework of the United Nations. Next, several topics are discussed that will require the attention of space lawyers in the near future. They are the International Space Station, space debris, exploitation of space resources, space tourism, private property rights in space, and militarization and weaponization of space. The paper raises some questions in each of these areas that need to be addressed and concludes that the general legal framework for space activities under public international law as contained in the UN treaties is in place, and is sufficiently general and flexible to enable and encourage states to carry out space activities in an orderly manner. However, as demonstrated by the examples discussed in the paper, the time has come for the international community to agree on the further development of these general principles, starting perhaps with space debris, imminent 'new' uses of space such as space tourism, or some of the 'age old' issues such as the weaponisation of outer space that will continue to require our attention and vigilance. Whether such rules can be in the form of non binding guidelines, codes of conduct and the like, or should be embodied in solid legal instruments creating rights and obligations remains to be seen.

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