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      • Prodomain Processing of Recombinant Plasmepsin II and IV, the Aspartic Proteases of Plasmodium falciparum, Is Auto- and Trans-Catalytic

        Man Kim, Yong,Hyang Lee, Mi,Gwang Piao, Tae,Wha Lee, John,Ho Kim, Jong,Lee, SungGa,Min Choi, Kyung,Hua Jiang, Jing,Ue Kim, Tae,Park, Hyun Japanese Biochemical Society 2006 The Journal of biochemistry Vol.139 No.2

        <P>Prodomain processing of the four food vacuole plasmepsins (PMs), the malarial aspartic proteases, is prerequisite for their activity on hemoglobin degradation of the parasite Plasmodium falciparum. Although previous studies have suggested the involvement of a calpain-like PM convertase in the processing of PMs, the underlying mechanism of their processing remains to be clarified. Here, to investigate the mechanism by which food vacuole PM II and IV are processed, we used their wild-type and mutant proteins in which the catalytic Asp residue in two active-site motifs was mutated, as well as protease inhibitors. Autocatalytic processing of wild-type PM II and IV was inhibited only by an aspartic protease inhibitor pepstatin A. Unexpectedly, their proteolytic activities were inhibited not only by pepstatin A but also by calpain inhibitor ALLN. The active-site mutants of both PM II and IV showed neither autocatalytic processing nor proteolytic activities. However, the mutants of both PMs were efficiently processed upon incubation with their respective wild type proteins. Furthermore, the mutants of both PMs were processed upon incubation with each other's wild-type PM in both pepstatin A- and ALLN-sensitive manners. These results suggest that the processing of PM II and IV occurs via an intra- and inter-molecular autocatalytic event as well as via a transcatalytic event between them.</P>

      • SCOPUSKCI등재

        살모넬라 C1 serogroup 특이 rfbM 유전자 증폭과 염기서열 분석

        이성일,정석찬,문진산,박용호,이존화,김병수,백병걸,Lee, Sung-il,Jung, Suk-chan,Moon, Jin-san,Park, Yong-ho,Lee, John-wha,Kim, Byeong-su,Baek, Byeong-kirl 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.1

        The Salmonella rfb gene encoding for the biosynthesis of the oligosaccharide-repeating units of the O-antigenic determinants was cloned and sequenced. A set of nucleotide primers(a forward and reverse) was selected to target a defined region of the guanosine diphospho-mannose(GDP-Man) pyrophosphorylase synthase gene : rfbM of Salmonella C serogroup. The primer set was used to develop a PCR-based rapid and specific detection system for Salmonella C1 serogroup. Amplification bands of predicted size(1,422bp) were generated from 11 different Salmonella C1 isolates. The bands were verified to be specific for the C1 serogroup by Southern blot analysis using reference homologous DNA specificity was further confirmed by the lack of reactivity with heterologous DNA derived from non-salmonella members of the family enterobacteriaeceae. A specificity of 100% was deduced along with a very high sensitivity shown by a detection limit of 1fg of a purified DNA template. The isolated DNA sequence was found to be 99.8% homologous to S montevideo but the related primers amplified with the predicted band sizes with all the Salmonella C1 serogroups tested. It is concluded that the PCR protocol based on the rfbM gene from S cholerasuis is optimal fast and specific for the detection of Salmonella C1 serogroup and also the corresponding probe is suitable for rapid detection of all Salmonella C1 serogroup DNA tested. This technology should facilitate the identification of contaminated pig products and for any other products contaminated with the Salmonalla C1 serogroup. The immediate impact of this developed method will be in the area of food safety of pig products with the potential prospect for adaptation to other food inspection technologies.

      • SCOPUSKCI등재

        소의 theileriosis가 성장 hormone과 insulin-like growth factor-I에 미치는 영향

        백병걸,변선윤,이존화,이호일,Baek, Byeong-kirl,Byoun, Sun-youn,Lee, John-wha,Lee, Ho-ill 대한수의학회 1997 大韓獸醫學會誌 Vol.37 No.2

        Bovine theileriosis caused by Theileria sergenti is the tick-borne intraery- throcytic piroplasmosis, that occurs in most regions of Korea. It results in severe economic losses on a farm caused by anemia, milk production loss, abortion and death. This study was undertaken to confirm the effects of the growth hormone and the insulin-like growth factor-I which are associated in the growth of cattle infected by T sergenti. The blood of one hundred and twenty ten-month Holstein was collected and the prepared blood smear was stained with acridine orange to investigate their parasitemia. And the hematological profiles were observed. According to the value of the hematocrit, they were categorized into four groups : Group 1 was under 20 percent, groups 2 and 3 were from over 21 to under 30 percent and from over 31 to under 35 percent and group 4 was over 36 percent. As the value of the hematocrit decreased, parasitemia(%) in erythrocytes was observed to increase(Y=-1.064X + 30.537, r=0.660). The amounts of the growth hormone and the insulin-like growth factor-I in the serum were measured by the radioimmunoassay. The growth hormone in serum of the group 1, group 2, group 3 and group 4 were observed as $0.238{\pm}0.043nmol/l$, $0.21{\pm}0.024nmol/l$, $0.366{\pm}0.035nmol/l$ and $0.646{\pm}0.223nmol/l$, respectively. The quantitative of the insulin-like growth factor-I in the same groups were observed also as $209.686{\pm}18.94ng/ml$, $250.9{\pm}12.609ng/ml$, $279.3{\pm}8.883ng/ml$ and $365.9{\pm}22.45ng/ml$, respectively. It can be concluded that the growth hormone and the insulin-like growth factor-I were observed to decrease in severe anemia due to theileriosis.

      • SCIESCOPUSKCI등재

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