• Korean Journal of Veterinary Research
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• v.42 no.3
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• pp.377-382
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• 2002

Bovine piroplasmosis caused by Theileria sergenti is a major cause of economic loss in livestock industry. Five cattle infected with Theileria sergenti showing severe and fatal anemia, confirmed by indirect immunofluorescent assay(IFA), were used in this study. Four cattle were treated with diminazene aceturate and one was not treated as the control. The therapeutic effect of diminazene aceturate against Theileria sergenti infection was monitored by detecting the 32kDa polypeptide specific for Theileria sergenti by the western blotting with both polyclonal and monoclonal antibodies. The 32kDa polypeptide detected at the beginning of diminazene aceturate treatment was not detectable after the treatment. It is postulated that the detection of the 32kDa polypeptide specific for Theileria sergenti may be a good tool for the diagnosis and monitoring the treatment progress of Theileria sergenti infection.

• Korean Journal of Veterinary Research
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• v.36 no.2
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• pp.453-461
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• 1996

Eighteen holstein-calves(4~5 months old) in a divided groups including the matched control were immunized with $100{\mu}g/dose$ of 34kDa, 45kDa polypeptide and T sergenti merozoite vaccine(protein content $100{\mu}g/dose$) respectively, previously mixed with aluminium hydroxide to elicit antibodies. All groups of calves were boosted with same dose and intervals. The animals were challenged by tick infestations in the endemic pasture of theileriosis from March to September 1994. The animals were monitored for the erythrocyte count, parasitemia, hematocrit and the specific antibody reactions elicited by immunization. The immunological responses demonstrated that vaccination with 34kDa polypeptide and T sergenti merozoite derived vaccine inhibited to produce the 75kDa band immunological responds even in the vaccinated calves after being challenged by tick infestations in the pasture. However, the specific antibody reactions were detected at the 32kDa band in the nonimmunized calves and T sergenti merozoite derived vaccine by the western blot. The 34kDa polypeptide vaccine and T sergenti merozoite derived vaccine were evaluated to be able to protect inducing anemia and to decrease parasitemias level. These vaccines have the efficacy of inhibition to produce a certain antigen corresponding 75kDa band antigen of parasite in the calves as challenged with tick infestations.

• Parasites, Hosts and Diseases
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• v.32 no.2
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• pp.111-116
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• 1994

Immunoblot analysis utilizing bovine sera from naturally or experimentally infected with Theileria sergenti were used to determine the immunodominant polypeptides of T sergenti (Korean isolated. The previously recognized major bands, 18 kDa,29 kDa, 34 kDa and 45 kDa, were excised after electrophoresis and transfer to PnF membrane. The individual bands were sequenced. The 34 kDa polypeptide which was the most antigenic and immunogenic peptide was observed in the Western blot. However, Chou-Fasman prediction sites (antigenic site) for antigen determinants of the 45 kDa, 34 kDa, 29 kDa and 18 kDa polypeptide were 6, 4, 2 and 0, respectively. However, the 45 kDa polypetide showed no reaction with anti- T sergenti hyperimmune serum.

• Journal of Plant Biology
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• v.39 no.4
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• pp.257-263
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• 1996

Flower-inducing activity in the phloem exudata of Pharbitis cotyledons was investigated using the bioassay of Pharbitis and Lemna. By SDS-PAGE and 2-D gel electrophoresis of the phloem exudate, two polypeptides of 11 kDa and of approximately 32 kDa (pI 6.9) showing qualitative changes during the flower induction were detected. A polypeptide of approximately 20 kDa (pI 4.8) specifically labeled in vivo with [35S]methionine was found during the inductive dark period in Pharbitis cotyledon tissues. The polypeptide of the equivalent molecular mass and with the identicl pI value was also detected by in vitro translation assay. Thus, it is assumed that the 20 kDa polypeptide plays a role in the process of flower induction in Pharbitis cotyledons.

• Journal of Pharmacopuncture
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• v.14 no.3
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• pp.71-79
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• 2011

Objectives : This study was undertaken to identify fibrinolytic proteases from Gloydius blomhoffii siniticus venom and to characterize a major fibrinolytic protease purified from the venom. Methods : The venom was subjected to chromatography using columns of Q-Sepharose and Sephadex G-75. The molecular weights of fibrinolytic proteases showing fibrinolytic zone in fibrin plate assay were determined in SDS-PAGE (Sodium dodecyl sulfate-polyacrylamide gel electrophoresis) The effects of inhibitors and metal ions on fibrinolytic protease and the proteolysis patterns of fibrinogen, gelatin, and bovine serum albumin were investigated. Results : 1) The fibrinolytic fractions of the three peaks isolated from Gloydius blomhoffii siniticus venom contained two polypeptides of 46 and 59 kDa and three polypeptides of 32, 18, and 15 kDa and a major polypeptide of 54 kDa, respectively. 2) The fibrinolytic activity of the purified protease of 54 kDA was inhibited by metal chelators, such as EDTA, EGTA, and 1,10-phenanthroline, and disulfhydryl-reducing compounds, such as dithiothreitol and cysteine. 3) Calcium chloride promoted the fibrinolytic activity of the protease, but mercuric chloride and cobalt(II) chloride inhibited it. 4) The fibrinolytic protease cleaved preferentially A${\alpha}$-chain and slowly B${\beta}$-chain of fibrinogen. It also hydrolyzed gelatin but not bovine serum albumin. Conclusions : The Gloydius blomhoffii siniticus venom contained more than three fibrinolytic proteases. The major fibrinolytic protease was a metalloprotease which hydrolyzed both fibrinogen and gelatin, but not bovine serum albumin.

• Korean Journal of Microbiology
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• v.32 no.3
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• pp.215-221
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• 1994

The complete nucleotide sequence of the cloned pga gene encoding the penicillin G acylase of Bacillus megaterium ATCC 14945 and its 5'- and 3'-flanking regions was determined. The sequence revealed only one large open reading frame (2,406 hp) of the penicillin G acylase (pga) gene. Upstream from ATG of the pga gene, there was a putative ribosome binding site, Shine-Dalgarno sequence. The promoter-like structure, - 10 and - 35 sequences, was also found. Following the stop codon, TAG, a structure reminiscent of the E. coli rho-independent transcription terminator was present. The amino acid sequence was deduced from the nucleotide sequence. The molecular mass of the polypeptide was 91,983 Da. There was a potential signal sequence in its amino-terminal region. A comparison of its deduced amino acid sequence with other characterized penicillin G acylases and the result of SDS-polyacrylamide gel electrophoresis of the purified enzyme showed that a precursor polypeptide of 92 kDa was processed into two dissimilar ${\alpha}$ and ${\beta}$-subunits of 25 and 61 kDa.

• BMB Reports
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• v.28 no.2
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• pp.138-142
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• 1995

An anticoagulant/fibrinolytic protease was purified to homogeneity from the earthworm Lumbricus rubellus. The protein was a single chain glycoprotein of 32 kDa that exhibited strong proteolytic activity on human thrombin and fibrin clots. Proteolytic degradation of these plasma proteins by the purified enzyme occurred at a neutral pH range. Among several human plasma proteins tested as possible substrates for the protease reaction, the 32 kDa enzyme specifically hydrolyzed both thrombin and fibrin polymers without affecting other proteins, such as serum albumin, immunoglobulin, and hemoglobin. Treatment of the purified enzyme at neutral pH with either phenylmethylsulfonylfluoride or soybean trypsin inhibitor resulted in a loss of catalytic activity. The enzyme hydrolyzed the chromogenic substrate H-D-Phe-L-Pipecolyl-L-Arg-p-nitroanilide with a $K_m$ value of 1.1 ${\mu}M$ at a neutral pH. These results suggest that the anticoagulant/fibrinolytic enzyme from Lumbricus rubellus is a member of the serine protease family having a trypsin-like active site, and one of the potential clevage sites for the enzyme is the carbonyl side of arginine residues in polypeptide chains.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.29 no.6
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• pp.797-804
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• 1996

Two trypsins were purified from shrimp hepatopancreas through ammonium sulfate fractionation, Q-Sepharose ionic exchange, benzamidine Sepharose-6B affinity, and Sephacryl S-300 gel chromatography. Both enzymes had a single polypeptide chain with a molecular weight (M.W.) of 32 kDa by sodium dodecylsulfate polyacrylamide gel electrophoresis (SOS-PAGE), although trypsin A and B were estimated to be a molecular weight of 27.2 and 22.8 kDa, respectively, using Sephacryl S-300 gel filtration. Both trypsins had similar amino acid compositions and rich in glycine, valine, alanine, aspartic acid, and glutamic acid, but low in methionine and basic amino acids. Both enzymes were completely inactivated by soybean trypsin inhibitor (SBTI), phenylmethylsulfonyl fluoride (PMSF), tosyl-L-lysine chloromethyl ketone (TLCK), benzamidine, leupeptin, however, not affected by tosyl-L-phenylalanine chloromethyl ketone (TPCK) and pepstatin.

• Animal cells and systems
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• v.13 no.3
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• pp.345-350
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• 2009

We have isolated and characterized a new insect chicken type (c-type) lysozyme gene from the common cutworm, Spodoptera litura. The full-length cDNA of Spodoptera lysozyme is cloned by rapid amplification of cDNA ends PCR (RACE-PCR). The isolated cDNA consists of 1039 bp including the coding region for a 142-amino acid residue polypeptide, which included a signal peptide of 21-amino acid residue and a mature protein of 121-amino acid residue. The predicted molecular weight of mature lysozyme and its theoretical isoelectric point from amino acid composition is 13964.8 Da and 9.05, respectively. The deduced amino acid sequence of Spodoptera lysozyme gene shows the highest similarity (96.7%) to Spodoptera exigua lysozyme among other lepidopteran species. Amino acid sequence comparison with other the c-type lysozymes, Spodoptera lysozyme has the completely conserved $Glu^{32}$ and $Asp^{50}$ of the active site and eight Cys residues are completely conserved in the same position as that of other lepidopteran lysozymes.

• Proceedings of the PSK Conference
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• 2003.04a
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• pp.300.1-300.1
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• 2003

Salmon calcitonin (sCT) is a therapeutic polypeptide hormone consisting of 32 amino acids (3432 Da). As with other bioactive peptide therapeutics, however, therapeutic use of sCT has been limited due to the problems of short circulating half-life and rapid proteolytic degradation. To get over this problem, the three positional isomers of mono-PEGylated sCT were prepared and among these, the best drug candiate for nasal application was chosen. (omitted)