• Proceedings of the Korean Society for Applied Microbiology Conference
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• 2002.10a
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• pp.18-25
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• 2002

The pikromycin biosynthetic system in Streptomyces venezuleae is unique for its ability to produce two groups of antibiotics that include the 12-membered ring macrolides methymycin and neomethymycin, and the 14-membered ring macrolides narbomycin and pikromycin. The metabolic pathway also contains two post polyketide-modification enzymes, a glycosyltransferase and P450 hydroxylase that have unusually broad substrate specificities. In order to explore further the substrate flexibility of these enzymes a series of hybrid polyketide synthases were constructed and their metabolic products characterized. The plasmid-based replacement of the multifunctional protein subunits of the pikromycin PKS in S. venezuelae by the corresponding subunits from heterologous modular PKSs resulted in recombinant strains that produce both 12- and 14-membered ring macrolactones with predicted structural alterations. In all cases, novel macrolactones were produced and further modified by the DesVII glycosyltransferase and PikC hydroxylase leading to biologically active macrolide structures. These results demonstrate that hybrid PKSs in S. venezuelae can produce a multiplicity of new macrolactones that are modified further by the highly flexible DesVII glycosyltransferase and PikC hydroxylase tailoring enzymes. This work demonstrates the unique capacity of the S. venezuelae pikromycin pathway to expand the toolbox of combinatorial biosynthesis and to accelerate the creation of novel biologically active natural products. The polyketide backbone of rifamycin B is assembled through successive condensation and ${\beta}$-carbonyl processing of the extender units by the modular rifamycin PKS. The eighth module, in the RifD protein, contains nonfunctional DH domain and functional KR domain, which specify the reduction of the ${\beta}$-carbonyl group resulting in the C-21 bydroxyl of rifamycin B. A four amino acid substitution and one amino acid deletion were introduced in the putative NADPH binding motif in the proposed KR domain encoded by rifD. This strategy of mutation was based on the amino acid sequences of the corresponding motif of the KR domain of module 3 in the RifA protein, which is believed dysfunctional, so as to introduce a minimum alteration and retain the reading frame intact, yet ensure loss of function. The resulting strain produces linear polyketides, from tetraketide to octaketide, which are also produced by a rifD disrupted mutant as a consequence of premature termination of polyketide assembly. Much of the structural diversity within the polyketide superfamily of natural products is due to the ability of PKSs to vary the reduction level of every other alternate carbon atom in the backbone. Thus, the ability to introduce heterologous reductive segments such as ketoreductase (KR), dehydratase (DH), and enoylreductase (ER) into modules that naturally lack these activities would increase the power of the combinatorial biosynthetic toolbox. The dehydratase domain of module 7 of the rifamycin PKS, which is predicted to be nonfunctional in view of the sequence of the apparent active site, was replaced with its functional homolog from module 7 of rapamycin-producing polyketide synthase. The resulting mutant strain behaved like a rifC disrupted mutant, i.e., it accumulated the heptaketide intermediate and its precursors. This result points out a major difficulty we have encountered with all the Amycolatopsis mediterranei strain containing hybrid polyketide synthases: all the engineered strains prepared so far accumulate a plethora of products derived from the polyketide chain assembly intermediates as major products instead of just analogs of rifamycin B or its ansamycin precursors.

• Journal of the Korea Institute of Information and Communication Engineering
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• v.10 no.1
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• pp.65-73
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• 2006

Network based VPN(Virtual Private Network) using MPLS(Multiprotocol Label Switching) technology, called PE(Provider Edge router) based MPLS VPN, is regarded as a good solution for intranets or ext3nets because of the low cost and the flexibility of the service provision. In this paper, we describe a mechanism that allows the VPN users to move from one site to another site of the VPN network based on the BGP-E MPLS technology. This mechanism is designed for PE(Provider Edge) routers of the backbone network. PE routers connected to the VPN sites establish a new MPLS path to the mobile node after they detect movement of the mobile VPN node. The new location may belong to the same VPN or to different VPN. We desisted VPN management and control functions of the PE routers in order to interface with the Mobile IP protocol and support the QoS mechanism. The pilot implementation and performance measurement were carried out on a simulation using COVERS tool.

• Journal of the Korean Magnetic Resonance Society
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• v.20 no.2
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• pp.56-60
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• 2016

Adenylate kinase (AK) enzyme which acts as the catalyst of reversible high energy phosphorylation reaction between ATP and AMP which associate with energetic metabolism and nucleic acid synthesis and signal transmission. This enzyme has three distinct domains: Core, AMP binding domain (AMPbd) and Lid domain (LID). The primary role of AMPbd and LID is associated with conformational changes due to flexibility of two domains. Three dimensional structure of human AK1 has not been confirmed and various mutation experiments have been done to determine the active sites. In this study, AK1R138A which is changed arginine[138] of LID domain with alanine[138] was made and conducted with NMR experiments, backbone dynamics analysis and mo-lecular docking dynamic simulation to find the cause of structural change and substrate binding site. Synthetic human muscle type adenylate kinase 1 (hAK1) and its mutant (AK1R138A) were re-combinded with E. coli and expressed in M9 cell. Expressed proteins were purified and finally gained at 0.520 mM hAK1 and 0.252 mM AK1R138A. Multinuclear multidimensional NMR experiments including HNCA, HN(CO)CA, were conducted for amino acid sequence analysis and signal assignments of $^1H-^{15}N$ HSQC spectrum. Our chemical shift perturbation data is shown LID domain residues and around alanine[138] and per-turbation value(0.22ppm) of valine[179] is consid-ered as inter-communication effect with LID domain and the structural change between hAK1 and AK1R138A.

• Bulletin of the Korean Chemical Society
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• v.27 no.4
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• pp.568-572
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• 2006

To study the effect of sequence on DNA structure, the two decamer crystal structures one alternating,d(GTACGCGTAC), and the other non-alternating, d(GGCCGCGGCC), were solved. Crystals of both decamers belong to the hexagonal space group $P6_122$, with one strand in the asymmetric unit. The unit cell constants of the alternating decamer are a = b = 39.26 $\AA$, c = 77.70 $\AA$. The structure was refined with 1,828 reflections from 8.0 to 2.0 Aresolution to an R value of 21.3% with all DNA atoms and 63 water molecules. The isomorphous non-alternating decamer had unit cell dimensions of a = b = 39.05 $\AA$, c = 82.15 $\AA$. The structure was refined with 2,423 reflections from 8.0 to 2.0 $\AA$ resolution to a final R value of 22.2% for all DNA atoms and 65 water molecules. Although the average helical parameters of the decamers are typical of A-DNAs, there are some minor differences between them. The helical twist, rise, x-displacement, inclination and roll alternate in the alternating decamer, but do not in the non-alternating decamer. The backbone conformations in both structures show some differences; the residue G(7) of the alternating decamer is trans for $\alpha$ and $\gamma$ while the trans conformations are observed at the residue G(8) of the non-alternating decamer.

• The Journal of Korean Institute of Communications and Information Sciences
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• v.28 no.10B
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• pp.927-937
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• 2003

Network based VPN(Virtual Private Network) using MPLS(Multiprotocol Label Switching) technology is regarded as a good solution for intranets or extranets because of the low cost and the flexibility of the service provision. In this paper, we describea mechanism that allows the VPN users to move from one site to another site of the VPN network based on the BGP-E MPLS technology. This mechanism is designed for and implemented on PE(Provider Edge) routers of the backbone network. PE routers connected to the VPN sites establish a new MPLS path to the mobile node after they detect movement of the mobile VPN node. The new location may belong to the same VPN or to different VPN. We designed VPN management and control functions of the PE routers in order to interface with the Mobile IP protocol. The pilot implementation and performance measurement were carried out on a testbed.

• Genomics & Informatics
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• v.14 no.2
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• pp.53-61
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• 2016

Toxoplasma gondii is an intracellular Apicomplexan parasite and a causative agent of toxoplasmosis in human. It causes encephalitis, uveitis, chorioretinitis, and congenital infection. T. gondii invades the host cell by forming a moving junction (MJ) complex. This complex formation is initiated by intermolecular interactions between the two secretory parasitic proteins-namely, apical membrane antigen 1 (AMA1) and rhoptry neck protein 2 (RON2) and is critically essential for the host invasion process. By this study, we propose two potential leads, NSC95522 and NSC179676 that can efficiently target the AMA1 hydrophobic cleft, which is a hotspot for targeting MJ complex formation. The proposed leads are the result of an exhaustive conformational search-based virtual screen with multilevel precision scoring of the docking affinities. These two compounds surpassed all the precision levels of docking and also the stringent post docking and cumulative molecular dynamics evaluations. Moreover, the backbone flexibility of hotspot residues in the hydrophobic cleft, which has been previously reported to be essential for accommodative binding of RON2 to AMA1, was also highly perturbed by these compounds. Furthermore, binding free energy calculations of these two compounds also revealed a significant affinity to AMA1. Machine learning approaches also predicted these two compounds to possess more relevant activities. Hence, these two leads, NSC95522 and NSC179676, may prove to be potential inhibitors targeting AMA1-RON2 complex formation towards combating toxoplasmosis.

• Journal of Korea Water Resources Association
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• v.51 no.4
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• pp.335-346
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• 2018

A vast amount of riverine spatial dataset have recently become available, which include hydrodynamic and morphological survey data by advanced instrumentations such as ADCP (Acoustic Doppler Current Profiler), transect measurements obtained through building various river basic plans, riverine environmental and ecological data, optical images using UAVs, river facilities like multi-purposed weir and hydrophilic sectors. In this regard, a standardized data model has been subsequently required in order to efficiently store, manage, and share riverine spatial dataset. Given that riverine spatial dataset such as river facility, transect measurement, time-varying observed data should be synthetically managed along specified river network, conventional data model showed a tendency to maintain them individually in a form of separate layer corresponding to each theme, which can miss their spatial relationship, thereby resulting in inefficiency to derive synthetic information. Moreover, the data model had to be significantly modified to ingest newly produced data and hampered efficient searches for specific conditions. To avoid such drawbacks for layer-based data model, this research proposed a relational data model in conjunction with river network which could be a backbone to relate additional spatial dataset such as flowline, river facility, transect measurement and surveyed dataset. The new data model contains flexibility to minimize changes of its structure when it deals with any multi-dimensional river data, and assigned reach code for multiple river segments delineated from a river. To realize the newly developed data model, Seom river was applied, where geographic informations related with national and local rivers are available.

• Journal of Legislation Research
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• no.44
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• pp.241-281
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• 2013

First and foremost, a discussion concerning government structure has to be done in connection with the state form and the governmental form. For practical reasons, there is a need to balance the principle of legality and its exceptions under the Government Organization Act. To ensure the flexibility of government structure with respect to the principle of legality, the National Assembly should accept the government structure requested by the newly elected government. This mitigates the rigidity of the principle of the legality within the government organizations. However, excessive changes by each government could violate the principle of legality asked by Constitution. In this sense, arbitrary modification with respect to the government structure by the newly elected government is not desirable. The long term stability of the government organization is required in any case. Secondly, general administrative agencies, other than Executive Ministries, should not be established under the direct order of the President without the control of the Prime Minister. A hierarchy of the executive branch (President->Prime Minister-> Executive Ministries) is stipulated in the Constitution. Establishing a hierarchy of President -> executive institution should be considered unconstitutional. Therefore, only the Presidential Secretariat and institutions with special functions can be established in the Presidential Office. Establishing general administrative agencies in the Presidential Office for convenience purposes is against the spirit of the current Constitution. Consequently, only the office of staffs and special agencies can be placed in the presidential office. It is against the spirit of the current Constitution to found administrative agencies under the presidential office for convenience. Thirdly, the office of the Prime Minister should be the backbone of internal affairs. In that sense, the President, as the head of state, should focus on the big picture such as the direction of the State, while the Cabinet headed by the Prime Minister should be responsible for the daily affairs of the State. The cabinet surrounding the Prime Minister must control all the ordinary affairs of the State, while the President, as the head of the State, should focus on the big picture of blueprinting the aim of the State. Lastly, the Office of the Prime Minister and Executive Ministries are the two main bodies of the executive branch. It is important to reduce the confusion caused by repeated changes in the names of Executive Ministries, to restore the traditional names and authorities of these institutions, and to rehabilitate the legitimacy of the State. For the Korean democracy to take its roots, a systematic way of stabilizing a law-governed democratic country is needed. There is also the need not only to reform security and economic agencies, but also to rationally solve the integration of technique and policy, according to the changes of time.