• Title/Summary/Keyword: Key site

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Characterization of the active site and coenzyme binding pocket of the monomeric UDP- galactose 4'- epimerase of Aeromonas hydrophila

  • Agarwal, Shivani;Mishra, Neeraj;Agarwal, Shivangi;Dixit, Aparna
    • BMB Reports
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    • v.43 no.6
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    • pp.419-426
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    • 2010
  • Aeromonas hydrophila is a bacterial pathogen that infects a large number of eukaryotes, including humans. The UDP-galactose 4'-epimerase (GalE) catalyzes interconversion of UDP-galactose to UDP-glucose and plays a key role in lipopolysaccharide biosynthesis. This makes it an important virulence determinant, and therefore a potential drug target. Our earlier studies revealed that unlike other GalEs, GalE of A. hydrophila exists as a monomer. This uniqueness necessitated elucidation of its structure and active site. Chemical modification of the 6xHis-rGalE demonstrated the role of histidine residue in catalysis and that it did not constitute the substrate binding pocket. Loss of the 6xHis-rGalE activity and coenzyme fluorescence with thiol modifying reagents established the role of two distinct vicinal thiols in catalysis. Chemical modification studies revealed arginine to be essential for catalysis. Site-directed mutagenesis indicated Tyr149 and Lys153 to be involved in catalysis. Use of glycerol as a cosolvent enhanced the GalE thermostability significantly.

The nonconserved N-terminus of protein phosphatases 1 influences its active site

  • Xie, XiuJie;Huang, Wei;Xue, ChengZhe;Wei, Qun
    • BMB Reports
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    • v.41 no.12
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    • pp.881-885
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    • 2008
  • Protein phosphatase 1 consists of a secondary structure arrangement, conserved in the serine/threonine protein phosphatase gene family, flanked by nonconserved N-terminal and C-terminal domains. The deletion mutant of PP1 with the 8 nonconserved N-terminal residues removed was designated PP1-(9-330). PP1-(9-330) had a higher activity and affinity than PP1 when assayed against four different substrates, and it also demonstrated a 6-fold higher sensitivity to the inhibitor okadaic acid. This suggested that the N-terminal domain suppresed the activity of PP1 and interfered with its inhibition by okadaic acid. The ANS fluorescence intensity of PP1-(9-330) was greater than that of PP1, which implies that the hydrophobic groove running from active site in the truncated PP1 was more hydrophobic than in PP1. Our findings provide evidence that the nonconserved N-terminus of PP1 functions as an important regulatory domain that influences the active site and its pertinent properties.

Analysis of Applicability of Visual SLAM for Indoor Positioning in the Building Construction Site (Visual SLAM의 건설현장 실내 측위 활용성 분석)

  • Kim, Taejin;Park, Jiwon;Lee, Byoungmin;Bae, Kangmin;Yoon, Sebeen;Kim, Taehoon
    • Proceedings of the Korean Institute of Building Construction Conference
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    • 2022.11a
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    • pp.47-48
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    • 2022
  • The positioning technology that measures the position of a person or object is a key technology to deal with the location of the real coordinate system or converge the real and virtual worlds, such as digital twins, augmented reality, virtual reality, and autonomous driving. In estimating the location of a person or object at an indoor construction site, there are restrictions that it is impossible to receive location information from the outside, the communication infrastructure is insufficient, and it is difficult to install additional devices. Therefore, this study tested the direct sparse odometry algorithm, one of the visual Simultaneous Localization and Mapping (vSLAM) that estimate the current location and surrounding map using only image information, at an indoor construction site and analyzed its applicability as an indoor positioning technology. As a result, it was found that it is possible to properly estimate the surrounding map and the current location even in the indoor construction site, which has relatively few feature points. The results of this study can be used as reference data for researchers related to indoor positioning technology for construction sites in the future.

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Damping modification factor of pseudo-acceleration spectrum considering influences of magnitude, distance and site conditions

  • Haizhong Zhang;Jia Deng;Yan-Gang Zhao
    • Earthquakes and Structures
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    • v.25 no.5
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    • pp.325-342
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    • 2023
  • The damping modification factor (DMF) is used to modify the 5%-damped response spectrum to produce spectral values that correspond to other necessary damping ratios for seismic design. The DMF has been the subject of numerous studies, and it has been discovered that seismological parameters like magnitude and distance can have an impact on it. However, DMF formulations incorporating these seismological parameters cannot be directly applied to seismic design because these parameters are not specified in the present seismic codes. The goal of this study is to develop a formulation for the DMF that can be directly applied in seismic design and that takes the effects of magnitude, distance, and site conditions into account. To achieve this goal, 16660 ground motions with magnitudes ranging from 4 to 9 and epicentral distances ranging from 10 to 200 km are used to systematically study the effects of magnitude, distance, and site conditions on the DMF. Furthermore, according to the knowledge that magnitude and distance affect the DMF primarily by changing the spectral shape, a spectral shape factor is adopted to reflect influences of magnitude and distance, and a new formulation for the DMF incorporating the spectral shape factor is developed. In comparison to the current formulations, the proposed formulation provides a more accurate prediction of the DMF and can be employed directly in seismic design.

Cloning and Characterization of Filamentous Fungal S-Nitrosoglutathione Reductase from Aspergillus nidulans

  • Zhou, Yao;Zhou, Shengmin;Yu, Haijun;Li, Jingyi;Xia, Yang;Li, Baoyi;Wang, Xiaoli;Wang, Ping
    • Journal of Microbiology and Biotechnology
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    • v.26 no.5
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    • pp.928-937
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    • 2016
  • S-Nitrosoglutathione reductase (GSNOR) metabolizes S-nitrosoglutathione (GSNO) and has been shown to play important roles in regulating cellular signaling and formulating host defense by modulating intracellular nitric oxide levels. The enzyme has been found in bacterial, yeast, mushroom, plant, and mammalian cells. However, to date, there is still no evidence of its occurrence in filamentous fungi. In this study, we cloned and investigated a GSNOR-like enzyme from the filamentous fungus Aspergillus nidulans. The enzyme occurred in native form as a homodimer and exhibited low thermal stability. GSNO was an ideal substrate for the enzyme. The apparent Km and kcat values were 0.55 mM and 34,100 min-1, respectively. Substrate binding sites and catalytic center amino acid residues based on those from known GSNORs were conserved in this enzyme, and the corresponding roles were verified using site-directed mutagenesis. Therefore, we demonstrated the presence of GSNOR in a filamentous fungus for the first time.

Thermostable Sites and Catalytic Characterization of Xylanase XYNB of Aspergillus niger SCTCC 400264

  • Li, Xin Ran;Xu, Hui;Xie, Jie;Yi, Qiao Fu;Li, Wei;Qiao, Dai Rong;Cao, Yi;Cao, Yu
    • Journal of Microbiology and Biotechnology
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    • v.24 no.4
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    • pp.483-488
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    • 2014
  • In order to improve the expression of heat-resistant xylanase XYNB from Aspergillus niger SCTCC 400264, XynB has been cloned into Pichia pastoris secretary vector pPIC9K. The XynB production of recombinant P. pastoris was four times that of E. coli, and the $V_{max}$ and specific activity of XynB reached $2,547.7{\mu}mol/mg$ and 4,757 U/mg, respectively. XynB still had 74% residual enzyme activity after 30 min of heat treatment at $80^{\circ}C$. From the van der Waals force analysis of XYNB (ACN89393 and AAS67299), there is one more oxygen radical in AAS67299 in their catalytic site, indicating that the local cavity is much more free, and it is more optimal for substrate binding, affinity reaction, and proton transfer, etc, and eventually increasing enzyme activity. The H-bonds analysis of XYNB indicated that there are two more H-bonds in the 33rd Ser of XYNB (AAS67299) than in the 33rd Ala(ACN89393 ), and two H-bonds between Ser70 and Asp67.

A Study on the Analysis of Risk Factors and the Reoccurrence Prevention in Construction Site Accidents (건설현장사고의 공종별 위험요소 분석 및 재발방지대책에 관한 연구)

  • Heo, Jun Kue;Choi, Mi Reu;Oh, Kwang Chin;Shin, Ju Yeoul
    • Journal of the Korea Institute of Construction Safety
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    • v.1 no.1
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    • pp.16-21
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    • 2018
  • The purpose of this study was to utilize 90 construction site accidents accumulated from 15 years ago by the Office of the High Level Investigation Committee of the Korea Infrastructure Safety Corporation under consignment from the Ministry of Land, Infrastructure and Transport. The construction site accident population used in this study is difficult to represent the entire construction site accident, but is considered to have statistical value as analysis data for a given size of construction site accident. The analysis is believed to be available on site as a basic data for securing construction safety and is expected to be used as a data for establishing future direction of construction safety policy.

The Expression of a Cytosolic Fructose-1,6-Bisphosphatase, a Key Enzyme in Sucrose Biosynthesis, Gene was Diurnally Fluctuated and Increased in Cold Acclimated Leaves of Chinese Cabbage

  • Leen, Jeong-Yeo;Song, Ha-Young;Lim, Yong-Pyo;Hur, Yoon-Kang
    • Journal of Plant Biotechnology
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    • v.33 no.2
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    • pp.123-131
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    • 2006
  • Chinese cabbage (Brassica rapa ssp. pekinesis) is one of the most important vegetable crops in korea and other East Asian countries. Cytosolic fructose-1,6-bisphospha-tase (cytFBPase) is a key enzyme in sucrose biosyn-thesis, which controls the sucrose levels as well as the productivity at plants. The Chinese cabbage cytFBPase gene, BrFBPase, encodes the 340 amino acid polypep-tide, giving a theoretical molecular weight of 37.2 kD and a isolectric point of 5.4. BrFBPase showed high sequence identity with Brassica homologs and its functional domains, such as 12,6P$_2$ binding site or active site and F6P binding site, were highly conserved in diverse sources of organisms. Although the genome of Chinese cabbage seemed to be triplicated, BrFBPase appears to be a single copy gene. The expression of BrFBPase was examined at transcript and protein levels under various conditions. BrFBPase expression was observed only in photosynthetic source tissue, not in sink tissue. The expression was slightly higher during the day than at night, and it showed a diurnal cycle with circadian rhythmicity. Short-term exposure to low temperature inhibited the expression of the BrFBPase, while long-term exposure increased the expression, supporting that sugar levels are high in late autumn when temperature are low.

Experimental study on seismic behavior of two-storey modular structure

  • Liu, Yang;Chen, Zhihua;Liu, Jiadi;Zhong, Xu
    • Steel and Composite Structures
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    • v.37 no.3
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    • pp.273-289
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    • 2020
  • Due to the unique construction method of modular steel buildings (MSBs) with units prefabricated fully off the site and assembled quickly on the site, the inter-module connection for easy operation and overall performance of the system were key issues. However, it was a lack of relevant research on the system-level performance of MSBs. This study investigated the seismic performance of two-storey modular steel structure with a proposed vertical rotary inter-module connection. Three full-scale quasi-static tests, with and without corrugated steel plate and its combination, were carried out to evaluate and compare their seismic behaviour. The hysteretic performance, skeleton curves, ductile performance, stiffness degradation, energy dissipation capacity, and deformation pattern were clarified. The results showed that good ductility and plastic deformation ability of such modular steel structures. Two lateral-force resistance mechanisms with different layout combinations were also discussed in detail. The corrugated steel plate could significantly improve the lateral stiffness and bearing capacity of the modular steel structure. The cooperative working mechanism of modules and inter-module connections was further analyzed. When the lateral stiffness of upper and lower modular structures was close, limited bending moment transfer may be considered for the inter-module connection. While a large lateral stiffness difference existed initially between the upper and lower structures, an obvious gap occurred at the inter-module connection, and this gap may significantly influence the bending moments transferred by the inter-module connections. Meanwhile, several design recommendations of inter-module connections were also given for the application of MSBs.

OBSERVATIONS OF EUV RECURRING JETS IN AN ACTIVE REGION CONFINED BY CORONAL LOOPS

  • Zheng, Yan-Fang;Wang, Feng;Ji, Kai Fan;Deng, Hui
    • Journal of The Korean Astronomical Society
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    • v.46 no.5
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    • pp.183-190
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    • 2013
  • Recurring jets, which are jets ejected from the same site, are a peculiar type among various solar jet phenomena. We report such recurring jets ejecting from the same site above an active region on January 22, 2012 with high-resolution multi-wavelength observations from Solar Dynamics Observatory(SDO). We found that the recurring jets had velocities, lengths and lifetimes, but had similar directions. The visible brightening appeared at the jet base before each jet erupted. All the plasma produced by the recurring jets could not overcome the large coronal loops. It seemed that the plasma ejecting from the jet base was confined and guided by preexisting coronal loops, but their directions were not along the paths of the loops. Two of the jets formed crossing structures with the same preexisting filament. We also examined the photospheric magnetic field at the jet base, and observed a visible flux emergence, convergence and cancellation. The four recurring jets all were associated with the impulsive cancellation between two opposite polarities occurring at the jet base during each eruption. In addition, we suggest that the fluxes, flowing out of the active region, might supply the energy for the recurring jets by examining the SDO/Helioseismic and Magnetic Imager (HMI) successive images. The observational results support the magnetic reconnection model of jets.