• Title/Summary/Keyword: Salt bridge

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Elucidation of Serpin's Conformational Switch Mechanism By Rapid Kinetic Study

  • Kang, Un-Beom;Lee, Cheolju;Baek, Je-Hyun;Seunghyun Ryu;Kim, Joon;Yu, Myeong-Hee
    • Proceedings of the Korean Biophysical Society Conference
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    • 2003.06a
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    • pp.62-62
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    • 2003
  • The native form of serpin (serine protease inhibitor) is kinetically trapped in metastable state. Metastability in these proteins is critical to their biological function. Serpins inhibit target proteases by forming a stable covalent complex in which the cleaved reactive site loop of the serpin is inserted into $\beta$-sheet A of the serpin with concomitant translocation of the protease to the opposite of the initial binding site. Despite recent determination of the crystal structures of a Michaelis protease-serpin complex as well as a stable covalent complex, details on the kinetic mechanism remain unsolved. In this study we constructed several $\alpha$$_1$-antitrypsin variants and examined their kinetic mechanism of loop translocation and formation of protease-serpin complex by stopped-flow experiments of fluorescence resonance energy transfer as well as quenched-flow experiment. We report here the relationship of serpin's conformational switch mechanism with Inhibitory activity. There is little direct correlation between loop insertion rate and inhibitory activity. Rather, disrupting a salt bridge between R196 and E354 accelerates loop translocation even though it impairs the inhibitory activity. Moreover, the serpin's reactive site loop is translocated, at least partially, prior to loop cleavage.

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Cryoscopy of Amine-Polytungstates (다중텅그스텐산 아민염의 분자량 측정)

  • Pyun, Chong-Hong;Sohn, Youn-Soo
    • Journal of the Korean Chemical Society
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    • v.18 no.2
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    • pp.126-131
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    • 1974
  • Trioctylamine-and tricaprylylmethylammonium chloride-tungstate salts have been prepared by solvent extraction from the sodium tungstate solution of various acidities(pH = 2, 4, 6). The molecular weights of the amine-tungstate salts thus obtained could be cryoscopically measured in benzene by means of a home-built Wheatstone bridge utilizing thermistor with sensitivity of 1/$4000^{\circ}C$. The cryoscopic data along with the results of chemical analysis and infrared spectra of the salts indicate that the amine-tungstates prepared at pH = 2 and 4 are all metatungstate whereas the salt obtained at pH = 6 is an unknown form quite different from the expected paratungstate.R = 0.14. By hydrogen bonding a guanidyl nitrogen of a sulfaguanidine molecule is linked to the sulfonyl oxygens of the other molecules indirectly through two different water molecules. The role of water molecule is both a .nor and an acceptor in hydrogen-bonding formation and these hydrogen bonds are tetrahedrally o?ented. The hydrogen-bonding networks form infinite molecular layers parallel to (001) plane.

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Three Non-Aspartate Amino Acid Mutations in the ComA Response Regulator Receiver Motif Severely Decrease Surfactin Production, Competence Development, and Spore Formation in Bacillus subtilis

  • Wang, Xiaoyu;Luo, Chuping;Liu, Youzhou;Nie, Yafeng;Liu, Yongfeng;Zhang, Rongsheng;Chen, Zhiyi
    • Journal of Microbiology and Biotechnology
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    • v.20 no.2
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    • pp.301-310
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    • 2010
  • Bacillus subtilis strains produce a broad spectrum of bioactive peptides. The lipopeptide surfactin belongs to one well-known class, which includes amphiphilic membrane-active biosurfactants and peptide antibiotics. Both the srfA promoter and the ComP-ComA signal transduction system are an important part of the factor that results in the production of surfactin. Bs-M49, obtained by means of low-energy ion implantation in wild-type Bs-916, produced significantly lower levels of surfactin, and had no obvious effects against R. solani. Occasionally, we found strain Bs-M49 decreased spore formation and the development of competence. Blast comparison of the sequences from Bs-916 and M49 indicate that there is no difference in the srfA operon promoter PsrfA, but there are differences in the coding sequence of the comA gene. These differences result in three missense mutations within the M49 ComA protein. RT-PCR analyses results showed that the expression levels of selected genes involved in competence and sporulation in both the wild-type Bs-916 and mutant M49 strains were significantly different. When we integrated the comA ORF into the chromosome of M49 at the amyE locus, M49 restored hemolytic activity and antifungal activity. Then, HPLC analyses results also showed the comA-complemented strain had a similar ability to produce surf actin with wild-type strain Bs-916. These data suggested that the mutation of three key amino acids in ComA greatly affected the biological activity of Bacillus subtilis. ComA protein 3D structure prediction and motif search prediction indicated that ComA has two obvious motifs common to response regulator proteins, which are the N-terminal response regulator receiver motif and the C-terminal helix-turn-helix motif. The three residues in the ComA N-terminal portion may be involved in phosphorylation activation mechanism. These structural prediction results implicate that three mutated residues in the ComA protein may play an important role in the formation of a salt-bridge to the phosphoryl group keeping active conformation to subsequent regulation of the expression of downstream genes.

Evaluation of the Properties of an Environment-Friendly De-icing Agent Based on Industrial By-Products (산업부산물을 활용한 친환경제설제의 특성평가)

  • Heo, Hyung-Seok;Lee, Byung-Jae
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.21 no.6
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    • pp.132-139
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    • 2017
  • A huge amount of de-icing agent is sprayed during winter to promote traffic safety in cold regions, and the quantity of de-icing agent sprayed has increased each year. The main ingredients in commonly used de-icing agents are chlorides, such as calcium chloride($CaCl_2$) and sodium chloride(NaCl). While calcium chloride is mostly used in Korea and sodium chloride is usually used in the U.S. and Japan, all de-icing agents include chloride ions. The chlorides included in sprayed calcium chloride-based de-icing agents have severe adverse effects, including the corrosion of reinforcing steels through salt damage by infiltrating into road structures, reduced structural performance of pavement or damage to bridge structures, and surface scaling, in combination with freezing damage in winter, as well as water pollution. In addition, the deterioration of paved concrete road surface that occurs after the use of calcium chloride-based de-icing agent accelerates the development of visual problems with traffic structures. Therefore, the present study was performed to prepare an environment-friendly liquid de-icing agent through a reaction between waste organic acids and calcium-based by-products, which are industrial by-products, and to analyze the properties of the de-icing agent in order to evaluate its applicability to road facilities.

Durability of concrete using sulfur-modified polymer (개질유황 폴리머를 사용한 콘크리트의 내구성 평가)

  • Hong, Chang Woo
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.25 no.5
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    • pp.205-211
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    • 2015
  • Most of the sulfur is obtained from desulfurization of natural gas and crude oil. In Korea, more than 120 tons of sulfur are produced by refinery, and about 50 % of the produced sulfur is used as a raw material for the production of fertilizer and sulfuric acid. Modified sulfur is manufactured from excessive sulfur that could be used to improve concrete properties, and this study evaluated concrete strength and durability that contains modified sulfur. Flexural and compressive strengths of concrete with sulfur modified polymer were comparable to those of OPC concrete with mixing water at similar temperatures, while the strengths increased a little as mixing water temperature increased. It was also confirmed that the resistance to freeze-thaw damage was more dependent on entrained air characteristics obtained by a proper use of air entraining agent than on the use of sulfur modified polymer. When concrete was immersed in 5 % sulfuric acid, the rate of reduction in compressive strength of OPC concrete was less than 1/4 of the strength reduction of concrete with sulfur modified polymer. Also, the resistance of concrete with sulfur modified polymer to scaling due to the use of de-icing salt was evaluated as Class 1, while that of OPC concrete was evaluated as Class 4, as aggregates were exposed. Accordingly, it is believed that sulfur modified polymer could be effectively used for bridge deck concrete since sulfur modified polymer improves the durability of concrete.

Prediction of Salinity of Nakdong River Estuary Using Deep Learning Algorithm (LSTM) for Time Series Analysis (시계열 분석 딥러닝 알고리즘을 적용한 낙동강 하굿둑 염분 예측)

  • Woo, Joung Woon;Kim, Yeon Joong;Yoon, Jong Sung
    • Journal of Korean Society of Coastal and Ocean Engineers
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    • v.34 no.4
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    • pp.128-134
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    • 2022
  • Nakdong river estuary is being operated with the goal of expanding the period of seawater inflow from this year to 2022 every month and creating a brackish water area within 15 km of the upstream of the river bank. In this study, the deep learning algorithm Long Short-Term Memory (LSTM) was applied to predict the salinity of the Nakdong Bridge (about 5 km upstream of the river bank) for the purpose of rapid decision making for the target brackish water zone and prevention of salt water damage. Input data were constructed to reflect the temporal and spatial characteristics of the Nakdong River estuary, such as the amount of discharge from Changnyeong and Hamanbo, and an optimal model was constructed in consideration of the hydraulic characteristics of the Nakdong River Estuary by changing the degree according to the sequence length. For prediction accuracy, statistical analysis was performed using the coefficient of determination (R-squred) and RMSE (root mean square error). When the sequence length was 12, the R-squred 0.997 and RMSE 0.122 were the highest, and the prior prediction time showed a high degree of R-squred 0.93 or more until the 12-hour interval.