• Title/Summary/Keyword: dimer formation

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Effects of the Heptasequence SPTSPTY of Rat Nuclear Factor 1-A on Interactions between the C-Terminal Regions of Mammalian Nuclear Factor 1 Proteins

  • Hwang, Jung-Su;Kim, Ji-Young
    • BMB Reports
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    • v.33 no.6
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    • pp.519-524
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    • 2000
  • NF1 proteins are a family of DNA binding proteins which consist of two separate domains, N-terminal DNA binding domain and C-terminal transcription activation domain. The N-terminal 220 amino acids are highly conserved and are also known to mediate dimerization of NF1 proteins. The C-terminal regions of different type of NF1 proteins are heterogeneous and responsible for transcriptional activation. In this study, we tested the interaction between different domains of rat NF1-A protein by yeast two hybrid analysis and observed the interaction between C-terminal regions of NF1-A which do not contain the N-terminal dimerization domain. Our results showed that the C-terminal region of rat NF1-A between residues 231 and 509 strongly interacted not only with itself, but also with human NF1/CTF1 which is a different type of NF1. When the C-terminal region was divided into two fragments, one from residue 231 to 447 and the other from 448 to 509, the two fragments were able to interact with the C-terminal region of NF1-A significantly. This indicates that both fragments contain independent interaction domains. Analysis of the interactions with alanine substituted fragments showed that substitutions of the heptasequence, SPTSPTY of NF1-A, affected interaction between NF1 proteins. Our results strongly suggest that C-terminal regions may also be important for the formation of homo- and heterodimers in addition to the N-terminal dimerization domain. Also, the heptasequence motif may play some roles in dimer formation.

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Behavior of AKD in AKD-sized Paper by Heating Treatment During Ageing (AKD 사이즈 처리한 종이의 저장중 가열처리에 의한 AKD의 거동)

  • Shin, Young-Doo;Seo, Won-Sung;Cho, Nam-Seok
    • Journal of Korea Technical Association of The Pulp and Paper Industry
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    • v.39 no.1 s.119
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    • pp.16-24
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    • 2007
  • This study was performed to investigate the reaction between alkylketene dimer(AKD) and cellulose molecules in AKD-sized paper sheet. AKD was added to highly beaten($80{\pm}3^{\circ}SR$) SwBKP(ca. 0.8% on pulp) in order to have much AKD retention in the paper sheet. This AKD-sized paper sheet was aged at different temperatures, $60^{\circ}C,\;80^{\circ}C,\;105^{\circ}C\;and\;125^{\circ}C$. Changes in FT-IR spectra of AKD in paper sheet during the ageing were measured. In addition, sizing degrees of the AKD-sized paper sheet pretreated for 30 sec. at $105^{\circ}C$ were measured by HST size tester during the storage at different temperature. IR spectra of AKD-sized paper sheet preheated at $105^{\circ}C$ for 30 sec. showed unchanged spectra two absorption bands at $1849cm^{-1}\;and\;1722cm^{-1}$ which refer to the typical AKD IR bands. However, these typical AKD bands were gradually reduced with increasing ageing, completely disappeared after 6 hrs. and formed new absorption band at $1706cm^{-1}$, which refers to carbonyl stretching vibration of dialkylktone. Eventually the AKD molecule was hydrolyzed to diakylketone without formation of ${\beta}$-ketoester with cellulose in paper sheet. After 6 days ageing, a little amount of ${\beta}$-ketoester bands was identified in 6 or 7 days ageing, because of the absence of water due to long-term heating. The same tendency was observed at different ageing conditions. At the practical papermaking process, AKD reacts prevailing with water, and mostly seems to be hydrolyzed to dialkylketene. Concerned to the sizing development, AKD-sized paper sheet was shown no sizing development at the initial stage of ageing at $60^{\circ}C$ after heating treatment at $105^{\circ}C$ for 30 sec., and gradually increased the sizing degree with increasing ageing, such as Hercules Sizing Tester (HST) 130 see for 12 hr, HST 300 sec. for 3 days and HST 400 sec. for 5 days. It was concluded that hydrolyzed AKD could contributed to the sizing development of the paper sheet.

Profiles of Local Fibrinolytic Activity before and after Urokinase Injection Into the Human Empyema Cavity (농흉환자에서의 늑막강내 유로키나제주입 전후의 섬유소 용해에 관한 연구)

  • Kim, Yong-Hoon;Kim, Jong-Bong;Moon, Jong-Ho;Song, Dong-Wha;Kim, Hyeon-Tae;Yang, Dong-Ho;Lee, Sang-Moo;Uh, Soo-Taek;Park, Choon-Sik
    • Tuberculosis and Respiratory Diseases
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    • v.40 no.4
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    • pp.378-383
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    • 1993
  • Background: In recent reports, it has been reported that increased coagulation and decreased fibrinolytic activity has been responsible for abnormal fibrin turnover in exudative pleural effusion. In the cases of empyema, the fibrinopurulent stage is characterized by the fibrin deposition resulting in formation of limiting membranes in the visceral and parietal pleura. Recently attention has been focused on intrapleural fibrinolytic therapy capable of removing intrapleural fibrin deposits by urokinase (UK) in the treatment of empyema. However, these clinical trials have provided the clinical evidences for resolution of pleural loculation after intrapleural urokinase injection (UK-injection), the profiles of fibrinolytic activity following the treatment were still not investigated. Therefore in order to demonstrate the fibrinolytic evidences behind the clinical efficacy of intracavitary UK-injection, we examined intrapleural plasminogen activator activity (PA-activity) and D-dimer (D-Di) concentrations before and after each repeated UK-injection into the pleura in subjects with loculated empyema cavity. Methods: In a group of 14 patients with multiple loculated empyema cavity, PA-activity and D-Di concentrations were measured before and after repeated UK-injection. One hundred thousand IU of UK was injected at each time and all sujects had at least two times of UK injection accoring to clinical decisions. Nine out of 14 sujects had three times of UK-injection. Results: The mean (${\pm}SE$) PA-activity prior to treatemnt was $10.5{\pm}7.0$ and it was increased to $91.9{\pm}27.0,\;432.3{\pm}177.1,\;170.0{\pm}85.3$ IU tPA/ml after first, second and third time of UK-injection respectively (p<0.01). D-Di concentrations were also increased from $4.16{\pm}1.06{\times}10^5$ to $9.62{\pm}1.54{\times}10^5,\;12.31{\pm}1.89{\times}10^5,\;8.54{\pm}1.56{\times}10^5$ ng/ml in the same order as above (p<0.05). Conclusion: The suppressed fibrinolytic activity in the empyema cavity get removed sinificantly after inrracavitary injection of urokinase by generation of additional intrapleural plasmin.

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Highly Efficient Synthesis of Conformationally Fixed Bicyclo[3.1.0]hexyl Nucleosides with an Ethenyl Group at C3'-Position as Potential Antiviral Agents

  • Kim, Seong Jin;Woo, Youngwoo;Park, Ah-Young;Kim, Hye Rim;Son, Sujin;Yun, Hwi Young;Chun, Pusoon;Moon, Hyung Ryong
    • Bulletin of the Korean Chemical Society
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    • v.35 no.9
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    • pp.2649-2654
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    • 2014
  • Synthesis of north-5'-methylbicyclo[3.1.0]hexyl adenine and hypoxanthine nucleosides with an ethenyl group at C3' position was successfully achieved by a highly facile method. Methylbicyclo[3.1.0]hexanone (${\pm}$)-7 with three contiguous chiral centers and its epimer (${\pm}$)-6 was remarkably simply constructed only by four steps involving a carbenoid insertion reaction in the presence of rhodium (II) acetate dimer as a metal catalyst, giving a correct relative stereochemistry of the generated three chiral centers. Due to steric hindrance from the concave face of the bicyclo[3.1.0]hexanone system, a Grignard reaction of (${\pm}$)-7 with ethenylmagnesium bromide showed exclusive diastereoselectivity towards the b-face. The Grignard reaction chemoselectively proceeded without reacting with ester functionality. Coupling reaction of glycosyl donor (${\pm}$)-11 with 6-chloropurine nucleobase afforded only the desired $N^9$-alkylated nucleoside without the formation of $N^7$-regioisomer. By the conventional method, 6-chloro group was converted into 6-amino and 6-hydroxy groups to give the desired adenine and hypoxanthine bicyclo[3.1.0]hexyl carbanucleosides with 3'-ethenyl group, respectively.

Dimerization of Aquooxomolybdenum (V) ion in Acid Media (I). Dehydrogenation of Bridging Hydroxide of $Mo_2O_4(OH)_{2(aq)}^{4+}$ (산성용액에서 아쿠오옥소몰리브덴 (V) 이온의 이합화 반응 (I). 이합체 착물의 두다리인 히드록소의 탈수소화 반응)

  • Chang-Su Kim;Chann-Woo Kim;Chang-Yong Kwon;Moon-Pyoung Yi
    • Journal of the Korean Chemical Society
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    • v.29 no.5
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    • pp.510-515
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    • 1985
  • Color of $MoO_{(aq)}^{3+}$ in concentrated methanesulfonic acid (∼10M) changes dark green due to the formation of $Mo_2O_2(OH)_{2(aq)}^{4+}$ dimer. This color is similar to that shown by addition of water to that shown by addition of water to green $MoO_{(aq)}^{3+}$ solution in 15-16M methanesulfonic acid. The molar extinction coefficient of monomer in 15M methanesulfonic acid is about 20 at 415nm. Rate constants are independent on the aquomolybdenum (V) and hydrogen ion concentration under the condition of this experment. Bridging hydroxides of $Mo_2O_2(OH)_{2(aq)}^{4+}$ are dehydrogenated at the less concentration of ∼6 M for HPTS and ∼10M for $CH_3SO_3H$. The structure of both the yl-oxygens and the bridging oxygens of final product is identified to (*image)unit.

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Protonation State of Norfloxacin and Their Interaction with DNA (pH에 따른 Norfloxacin의 형태 및 DNA와의 상호작용에 관한 연구)

  • Yeo, Jeong-Ah;Son, Gwan-Su;Kim, Jong-Moon;Moon, Hyung-Rang;Jun, Eun-Duk;Cho, Tae-Sub
    • Journal of the Korean Chemical Society
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    • v.44 no.1
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    • pp.4-9
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    • 2000
  • We investigated the self-association and protonation state of norfloxacin, a member of quinokme antibiotics, using electric absorption and potentiom,etric titration. Both nitrogen at the piperazine ring and carboxylic acid were protonated at a low pH (cationic norfloxacin), and deprotonated at a high pH (anionic norfloxacin). In the intermediate pH range, a neutral species was dominant with the possibility of forming a zwitter ion. We also observed that nortloxacin molecules can be sracked to form a dimer at an intermediate pH, The equilibrium constant of the norfloxacin-DNA complex formation, which was measured by Stem-Volmermethod, increases as the pH of the system is lowered. This observation indicates that it is the cationic nortloxacin that forms a complex with DNA among various norfloxacin species in aqueous solution.

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Ligand Binding Properties of the N-Terminal Domain of Riboflavin Synthase from Escherichia coli

  • Lee, Chan-Yong;Illarionov, Boris;Woo, Young-Eun;Kemter, Kristina;Kim, Ryu-Ryun;Eberhardt, Sabine;Cushman, Mark;Eisenreich, Wolfgang;Fischer, Markus;Bacher, Adelbert
    • BMB Reports
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    • v.40 no.2
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    • pp.239-246
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    • 2007
  • Riboflavin synthase from Escherichia coli is a homotrimer of 23.4 kDa subunits and catalyzes the formation of one molecule each of riboflavin and 5-amino-6-ribitylamino- 2,4(1H,3H)-pyrimidinedione by the transfer of a 4-carbon moiety between two molecules of the substrate, 6,7- dimethyl-8-ribityllumazine. Each subunit comprises two closely similar folding domains. Recombinant expression of the N-terminal domain is known to provide a $C_2$-symmetric homodimer. In this study, the binding properties of wild type as well as two mutated proteins of N-terminal domain of riboflavin synthase with various ligands were tested. The replacement of the amino acid residue A43, located in the second shell of riboflavin synthase active center, in the recombinant N-terminal domain dimer reduces the affinity for 6,7-dimethyl-8-ribityllumazine. The mutation of the amino acid residue C48 forming part of activity cavity of the enzyme causes significant $^{19}F$ NMR chemical shift modulation of trifluoromethyl derivatives of 6,7-dimethyl-8-ribityllumazine in complex with the protein, while substitution of A43 results in smaller chemical shift changes.

Disulfide Bond Bridged Divalent Antibody-Toxin, $(Fab-PE38fl)_2$ with the Toxin PE38 Fused to the Light Chain

  • Won, Jae-Seon;Choe, Mu-Hyeon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.8
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    • pp.1475-1481
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    • 2008
  • B3 antibody specifically binds the $Lewis^Y$-related carbohydrate antigen of many carcinomas, and it is used as a model antibody in this study. In a previous study, the Fab fragment of the antibody was fused to a 38 kDa truncated form of Pseudomonas exotoxin A, PE38, to make Fab-PE38, where PE38 is fused to the Fd fragment of the Fab domain. This parent monomer molecule, Fab-PE38, had no cysteine in the hinge region, and it could not make a disulfide bond to form a disulfide bond bridged homodimer. In this study, we constructed three different kinds of divalent Fab-toxin fusion homodimers where the toxin is fused to the light chain of Fab, $(Fab-PE38fl)_2$. In addition to the PE38 toxin fused to the light chain, these three molecules have different hinge sequences hi, h2, and h3 making Fabh1-, Fabh2-, and Fabh3-PE38fl monomers, respectively. These hinges contain only one cysteine on different positions of the hinge sequence. The disulfide bond between the hinge region of two monomers forms homodimers $(Fabh1-PE38fl)_2$, $(Fabh2-PE38fl)_2$, and $(Fabh3-PE38fl)_2$. The refolding yields of these dimers were 5-16-fold higher than a previously constructed dimer where the PE38 was fused to the Fd fragment $(Fabh2-PE38)_2$ [8]. Our data suggest that the steric repulsion between the two PE38s in $(Fabh1-PE38)_2$ during disulfide bridge formation is relieved by fusing it at the end of the light chain. The best cytotoxicity value of these dimers showed about 2.5-fold higher on an MCF7 cell line than that of the monovalent reference molecule in ng/ml scale, which is 15-fold higher in pM scale.

Evaluation of the in vivo Antithrombotic, Anticoagulant and Fibrinolytic Activities of Lumbricus rubellus Earthworm Powder

  • Hahn, Bum-Soo;Jo, You-Young;Yang, Kyung-Youl;Wu, Song-Ji;Pyo, Mi-Kyung;Yunchoi, Hye-Sook;Kim, Yeong-Shik
    • Archives of Pharmacal Research
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    • v.20 no.1
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    • pp.17-23
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    • 1997
  • A saline suspension of Lumbricus rubellus earthworm powder (EWP) was administered to rats (1 g/kg/day) orally for 15 days to evaluate an oral effectiveness for thrombotic disorders. Blood was drawn at 2-day interval after the administration. Several parameters for antithrombotic, anticoagulant and fibrinolytic activities were measured, including platelet aggregation, clotting time, plasmin activity and the levels of FDP (fibrin/fibrinogen degradation products), D-dimer, and t-PA antigen. It did not affect platelet aggregation induced by ADP and collagen but anticoagulant activity (aPTT and TT) was gradually increased to two-folds for the first 5 days of administration and back to normal. Fibrinolytic activity of euglobulin fraction was highest on the 11 th day after the administration. The level of FDP was elevated to be comparable to the positve control$ (5-10 {\mu}g/ml)$ after 9-day treatment. Oral administration of the EWP could also reduce the formation of venous thrombus induced with viper venom. Complete blood count (CBC) profiles were within normal ranges except for a slight increase in white blood cells after the oral administration for 15 days. These results suggested that the EWP may be valuable for the prevention and/or treatment of thrombotic diseases.

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Reaction Kinetic Study on Pyrolysis of Waste Polystyrene using Wetted Column Reactor (Wetted Column 반응기를 이용한 폴리스티렌 열분해 반응속도론적 연구)

  • You, Young Gil;Yoon, Byung Tae;Kim, Seong Bo;Choi, Myoung Jae;Choi, Cheong Song
    • Korean Chemical Engineering Research
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    • v.46 no.3
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    • pp.535-539
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    • 2008
  • Conversion to oil, yield of styrene and formation of side products such as ${\alpha}-methyl$ styrene, ethyl benzene, benzene, toluene, dimer and trimer were affected by residue formed during thermal degradation. Also, control of reaction temperature had a difficulty at the first stage. Thus, new reaction system using wetted-wall type reactor was proposed and examined on various parameters such as reaction temperature, feeding rate and removal velocity of formed vapor. Optimun condition was obtained from continuous thermal degradation using wetted-wall type reactor and reaction kinetic study was carried out at new type reactor.