• Title/Summary/Keyword: Barrel Assembly

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Comparative Analysis of Envelope Proteomes in Escherichia coli B and K-12 Strains

  • Han, Mee-Jung;Lee, Sang-Yup;Hong, Soon-Ho
    • Journal of Microbiology and Biotechnology
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    • v.22 no.4
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    • pp.470-478
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    • 2012
  • Recent genome comparisons of E. coli B and K-12 strains have indicated that the makeup of the cell envelopes in these two strains is quite different. Therefore, we analyzed and compared the envelope proteomes of E. coli BL21(DE3) and MG1655. A total of 165 protein spots, including 62 nonredundant proteins, were unambiguously identified by two-dimensional gel electrophoresis and mass spectrometry. Of these, 43 proteins were conserved between the two strains, whereas 4 and 16 strain-specific proteins were identified only in E. coli BL21(DE3) and MG1655, respectively. Additionally, 24 proteins showed more than 2-fold differences in intensities between the B and K-12 strains. The reference envelope proteome maps showed that E. coli envelope mainly contained channel proteins and lipoproteins. Interesting proteomic observations between the two strains were as follows: (i) B produced more OmpF porin with a larger pore size than K-12, indicating an increase in the membrane permeability; (ii) B produced higher amounts of lipoproteins, which facilitates the assembly of outer membrane ${\beta}$-barrel proteins; and (iii) motility- (FliC) and chemotaxis-related proteins (CheA and CheW) were detected only in K-12, which showed that E. coli B is restricted with regard to migration under unfavorable conditions. These differences may influence the permeability and integrity of the cell envelope, showing that E. coli B may be more susceptible than K-12 to certain stress conditions. Thus, these findings suggest that E. coli K-12 and its derivatives will be more favorable strains in certain biotechnological applications, such as cell surface display or membrane engineering studies.

Structure of the Tripartite Multidrug Efflux Pump AcrAB-TolC Suggests an Alternative Assembly Mode

  • Kim, Jin-Sik;Jeong, Hyeongseop;Song, Saemee;Kim, Hye-Yeon;Lee, Kangseok;Hyun, Jaekyung;Ha, Nam-Chul
    • Molecules and Cells
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    • v.38 no.2
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    • pp.180-186
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    • 2015
  • Escherichia coli AcrAB-TolC is a multidrug efflux pump that expels a wide range of toxic substrates. The dynamic nature of the binding or low affinity between the components has impeded elucidation of how the three components assemble in the functional state. Here, we created fusion proteins composed of AcrB, a transmembrane linker, and two copies of AcrA. The fusion protein exhibited acridine pumping activity, suggesting that the protein reflects the functional structure in vivo. To discern the assembling mode with TolC, the AcrBA fusion protein was incubated with TolC or a chimeric protein containing the TolC aperture tip region. Three-dimensional structures of the complex proteins were determined through transmission electron microscopy. The overall structure exemplifies the adaptor bridging model, wherein the funnel-like AcrA hexamer forms an intermeshing cogwheel interaction with the ${\alpha}$-barrel tip region of TolC, and a direct interaction between AcrB and TolC is not allowed. These observations provide a structural blueprint for understanding multidrug resistance in pathogenic Gram-negative bacteria.

Reliability Analysis Using Field-Data of 5.56 mm Rifle (야전운용제원을 이용한 5.56 mm 소총 신뢰도 분석)

  • Shin, Tae-Sung;Seo, Hyun-Soo;Lee, Ho-Jun;Choi, Si-Young;Gil, Hyeon-Jun
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.21 no.6
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    • pp.584-591
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    • 2020
  • Reliability is an important factor in weapons systems. Low reliability causes the weapons system to fail to function properly, which directly leads to the weakening of combat capability. This paper classifies the structure of the 5.56 mm rifle, which is currently used by the Korean army, into a total of seven assemblies and describes the eight functions necessary for the rifle to operate normally. In addition, the concept of reliability was defined as the MRBF, and the Poisson process model and TTT plot were explained as a reliability analysis theory for the repair function system. Next, the field-data obtained by defining failure as the replacement of parts other than periodic exchange of parts were refined, and the reliability was analyzed by entering the refined field operation specifications into the Minitab program. As a result, the reliability of the rifle was determined to be 251.73. The assembly parts that required improvement was identified as the barrel, lower body, and butt stock assembly, and 10 detailed parts needed to be improved. Finally, the limits of the reliability analysis using the field-data currently available for small caliber firearms were considered.

Dynamic Behavior of Reactor Internals under Safe Shutdown Earthquake (안전정기지진하의 원자로내부구조물 거동분석)

  • 김일곤
    • Computational Structural Engineering
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    • v.7 no.3
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    • pp.95-103
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    • 1994
  • The safety related components in the nuclear power plant should be designed to withstand the seismic load. Among these components the integrity of reactor internals under earthquake load is important in stand points of safety and economics, because these are classified to Seismic Class I components. So far the modelling methods of reactor internals have been investigated by many authors. In this paper, the dynamic behaviour of reactor internals of Yong Gwang 1&2 nuclear power plants under SSE(Safe Shutdown Earthquake) load is analyzed by using of the simpled Global Beam Model. For this, as a first step, the characteristic analysis of reactor internal components are performed by using of the finite element code ANSYS. And the Global Beam Model for reactor internals which includes beam elements, nonlinear impact springs which have gaps in upper and lower positions, and hydrodynamical couplings which simulate the fluid-filled cylinders of reactor vessel and core barrel structures is established. And for the exciting external force the response spectrum which is applied to reactor support is converted to the time history input. With this excitation and the model the dynamic behaviour of reactor internals is obtained. As the results, the structural integrity of reactor internal components under seismic excitation is verified and the input for the detailed duel assembly series model could be obtained. And the simplicity and effectiveness of Global Beam Model and the economics of the explicit Runge-Kutta-Gills algorithm in impact problem of high frequency interface components are confirmed.

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