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Mutational Analysis of Two Conserved Active Site Tyrosine Residues in Matrilysin

  • Jaeho Cha
    • Journal of Life Science
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    • v.9 no.2
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    • pp.44-48
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    • 1999
  • The ionization of tyrosine residue is known to be involved in the stabilization of transition-state in catalysis of astacin based upon the astacin-transition state analog structure. Two tyrosine residues, Tyr-216 and Tyr-219, are conserved in all MMPs related with astacin family, We replaced Tyr-216 and Tyr-219 into phenylalanine, respectively and the zinc binding properties, kinetic parameters, and pH dependence of each mutant are determined in order to examine the role of tyrosine residue in matrilysin catalysis. Both mutants contain two zinc atoms per mol of enzyme, indicating that either tyrosime does not affect the zinc binding property of the enzyme. Y216F and Y219F mutants are highly active and the kcat/Km values are only decreased 1.1-1.5-fold compared to the wild-type enzyme. The decrease in the activity of the mutants is essentially due to the increase in Km value. The pH dependencies of the kcat/Km values for both mutants are similar to the corresponding dependencies obtained with the wild type enzyme. The pKa values at the alkaline side of both mutants are not changed. These kinetic and pH dependence results indicate that the ionization of active site tyrosine residue of matrilysin is not reflected in the kinetics of peptide hydrolysin as catalyzed by astacin.

Rapid Screen for Bacteria Solubilzing Insoluble Phoshpate on Agar Plate

  • Son, Hong-Joo;Kang, Sung-Il;Kim, Yong-Gyun;Kim, Hee-Goo;Lee, Sang-Joon
    • Journal of Life Science
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    • v.10 no.1
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    • pp.64-65
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    • 2000
  • Insoluble phosphate-solubilizing bacteria are routinely screened by a plate assay method using Pikovskaya agar and a modified Pikovskaya medium. A modified Pikovskaya medium to improve the clarity of the yellow-colored halo has not necessarity improved the plate assay. Colonies of phosphate-solubilizing bacteria tested could be redily selected after 48 h of incubation by green-colored colony formation on plate in which bromcresol green(BCG) was included. Among them, two bacterial strains did not produce distinct yellow halos after 48 h of incubation. We suggest that the green colony formation on plate medium containing BCG is advantageous ofr rapid isolating phosphate-solubilizing bacteria directly from soil.

Cytogenetic Studies in Hybrids from a Pair of Sibling Drosophila Species

  • Park, Yung-Hyun;Kim, Heui-Soo;Lee, Won-Ho
    • Journal of Life Science
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    • v.10 no.1
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    • pp.48-50
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    • 2000
  • The cytogenetic pattern of autosome and sex chromiosome after Giemas staining were examined in the hybrids between two sibling species, Drosophila melanogaster and D. simulans. The analysis of karyotype in the hybrid female between D. melanogaster females and D. simulans males could be easily distinguished the characteriation of eight chromosomes from bothe species, especially with regard to X chromosomes. The lagging duplication of Y chromosome was investigated in the interspecific hybrid males from the cross between female of Drosophila melanogaster(OR) and males of D. simulasn (K18). On the other hand, the X chromatids of D. simulans were loosely separated in the early stage of anaphase.

Evidence for a Catalytic Role of Glutamic Acid 233 of Yac-1 in Arginine-Specific ADP-Ribosylation of Murine Lymphocyte

  • Kim, Hyun-Ju
    • Journal of Life Science
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    • v.10 no.1
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    • pp.20-23
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    • 2000
  • Mono-ADP-ribosylation, catalyzed by ADP-ribosyltransferases, is a post-translational modification of proteins in which the ADP-ribose moiety of NAD is transferred to an acceptor protein. Previously, we have identified and cloned a glycosylphosphatidylinositol-linked ADP-ribosyltransferase (Yac-1) from mouse lymphoma cells. Yac-1 enzyme contains three regions (region I,II,III) similar to those found in several bacterial toxins and vertebrate ADP-ribosyltransferases. Site-directed mutagenesis was performed to verify the role of Glu 233 in region III. Mutants E233Q, E233D and E233A were inactive for ADP-ribosyltransferase activity. Thus Glu 233 in Yac-1 is essential for enzyme activity, suggesting that Glu 233 in Glu-rich motif near the carboxy terminus plays a catalytic role in ADP-ribosyltransferase activity.

Identification of the Invasion Determinants of Salmonella typhimurium for Cultured HEp-2 and HeLa Cells

  • Park, Jeong-Uck;Joo, Woo-Hong
    • Journal of Life Science
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    • v.10 no.1
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    • pp.6-9
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    • 2000
  • Salmonella typhimurium is a causative agent of the common worldwide disease, salmonellosis. To identify putative invasion genes involved in Samonella infections, a S. typhimurium cosmid library was constructed in noninvasive E. coli DJl. The invasion efficiencies of the cosmid library for cultured HEp-2 and HeLa cells were estimated by tissue-culture invasion assay. 2 out of 1,000 transductants, DHl(pSI623) and DHl(pSI511) were able to invade the cells. Compared to E. coli by DHl(pSI511) increased 25- and 33 fold, respectively. The invasion efficiencies of HeLa cells by DHl(pSI623) increased 31- and 35 fold, respectively. This illustrates that the cosmid clones, DHl(pSI623) and DHl(pSI511) could harbor the invasion determinants derived from genomic DNA of S. typhimurium 82/6915, conferring the invasive characters for the cells.

STRONG CONVERGENCE IN NOOR-TYPE ITERATIVE SCHEMES IN CONVEX CONE METRIC SPACES

  • LEE, BYUNG-SOO
    • The Pure and Applied Mathematics
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    • v.22 no.2
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    • pp.185-197
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    • 2015
  • The author considers a Noor-type iterative scheme to approximate com- mon fixed points of an infinite family of uniformly quasi-sup(fn)-Lipschitzian map- pings and an infinite family of gn-expansive mappings in convex cone metric spaces. His results generalize, improve and unify some corresponding results in convex met- ric spaces [1, 3, 9, 16, 18, 19] and convex cone metric spaces [8].

A Post-Analysis of Decision Tree to Detect the Change of Customer Behavior on Internet Shopping Mall

  • Kim, Jae kyeong;Song, Hee-Seok;Kim, Tae-Sung
    • Proceedings of the Korea Inteligent Information System Society Conference
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    • 2001.01a
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    • pp.456-463
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    • 2001
  • Understanding and adapting to changes of customer behavior in internet shopping mall is an important aspect to survive in continuously changing environment. This paper develops a methodology based on decision tree algorithms to detect changes of customer behavior automatically from customer profiles and sales data at different time snapshots. We first define three types of changes as emerging pattern, unexpected change and the added/perished rule. Then, it is developed similarity and difference measures for rule matching to detect all types of change. Finally, the degree of change is developed to evaluate the amount of change. A Korean internet shopping mall case is evaluated to represent the performance of our methodology. And practical business implications for this methodology are also provided.

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Spatial Query Processing Based on Minimum Bounding in Wireless Sensor Networks

  • Yang, Sun-Ok;Kim, Sung-Suk
    • Journal of Information Processing Systems
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    • v.5 no.4
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    • pp.229-236
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    • 2009
  • Sensors are deployed to gather physical, environmental data in sensor networks. Depending on scenarios, it is often assumed that it is difficult for batteries to be recharged or exchanged in sensors. Thus, sensors should be able to process users' queries in an energy-efficient manner. This paper proposes a spatial query processing scheme- Minimum Bounding Area Based Scheme. This scheme has a purpose to decrease the number of outgoing messages during query processing. To do that, each sensor has to maintain some partial information locally about the locations of descendent nodes. In the initial setup phase, the routing path is established. Each child node delivers to its parent node the location information including itself and all of its descendent nodes. A parent node has to maintain several minimum bounding boxes per child node. This scheme can reduce unnecessary message propagations for query processing. Finally, the experimental results show the effectiveness of the proposed scheme.

Microbiological Identification of Medical Probiotic Bisspan Strain (의약용 프로바이오틱 비스판균의 미생물학적 동정)

  • 전경동;이광호;김원석;백현동
    • Microbiology and Biotechnology Letters
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    • v.28 no.2
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    • pp.124-127
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    • 2000
  • Beneficial bacteria, which have been used for medical purpose and for medicines for treating intestinal disorders, include strains of Bifidobacterium sp., Lactobacillus sp., Enterococcus sp., Clostridium butyricum, Lactobacillus sporogenes, Bacillus subtilis, Bacillus polyfermenticus and the like. Bacillus polyfermenticuss SCD with is commonly called as Bispan strain has been appropriately used for the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine. In this study, the identification and characterization of Bispan strain was done using SEM observation, API 50CHB kits, isoprenoid quinone analysis, and fatty acid analysis. These results suggest that Bispan strain is very similar to Bacillus subtilis.

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Genus Diversity of Soil Actinomycetes Isolated from Natural Lime Cave. (자연 석회동굴에서 분리한 방선균의 속 다양성)

  • 박동진;이상화;박해룡;권오성;박상호;마사카즈우라모토;김창진
    • Microbiology and Biotechnology Letters
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    • v.28 no.3
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    • pp.129-133
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    • 2000
  • Different actinomycete strains were isolated from natural lime caves of Ondal Chemongok Hwanseon and Yongyeon which are located at Kangwon or chungcheongbook province in Korea and were identified to the genus level. Soil sam-ples were collected at 6 sites inside and 2 sites outside of each natural lime cave, As the result the strains belonging to genus Streptomyces and rare actinomycetes were isolated at the average of 2.1 and 3.4 strains per g soil on inside cave whereas which were isolated at the 6.0 and 1.8 strains per g soil on outside cave. How-ever the generic distribution of Streptomyces and rare actinomycetes isolated from outside cave was quite dif-ferent from that of inside cave. It was shown that rare actinomycetes at natural lime caves is generally highly abundant than Streptomyces.

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