• Journal of Pharmacopuncture
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• v.17 no.1
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• pp.44-50
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• 2014

Objectives: This study was undertaken to isolate a fibrinolytic enzyme from the snake venom of Gloydius blomhoffii siniticus and to investigate its enzymatic characteristics and hemorrhagic activity as a potential pharmacopuncture agent. Methods: The fibrinolytic enzyme was isolated by using chromatography, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and fibrin plate assay. The characteristics of the enzyme were investigated using fibrin plate assay, protein hydrolysis analysis, and hemorrhage assay. Its amino acid composition was determined. Results: The fibrinolytic enzyme with the molecular weight of 32kDa (FE-32kDa) from Gloydius blomhoffii siniticus showed a fibrin hydrolysis zone at the concentration of 0.2 mg/mL in the fibrin plate assay. The fibrin hydrolysis activity of the enzyme was inhibited completely by ethylenediaminetetraacetic acid (EDTA), ethyleneglycoltetraacetic acid (EGTA), and 1, 10-phenanthroline, thiothreitol and cysteine, and partially by phenylmethanesulfonylfluoride (PMSF). Metal ions such as $Fe^{2+}$ and $Hg^{2+}$ inhibited the fibrin hydrolysis completely, but $Zn^{2+}$ enhanced it. FE-32kDa hydrolyzed ${\alpha}$-chain but did not hydrolyze ${\beta}$-chain and ${\gamma}$-chain of fibrinogen. High-molecular-weight polypeptides of gelatin were hydrolyzed partially into low-molecular-weight polypeptides, but the extent of hydrolysis was limited. FE-32kDa induced hemorrhage beneath back skin of mice at the dose of $2{\mu}g$. Conclusions: FE-32kDa is a ${\alpha}$-fibrin(ogen)olytic metalloprotease that requires $Zn^{2+}$ for fibrinolytic activity and causes hemorrhage, suggesting that the enzyme is not appropriate for use as a clinical pharmacopuncture.

• International Journal of Industrial Entomology and Biomaterials
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• v.2 no.2
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• pp.167-172
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• 2001

The vitellin of firefly, Pyrocoelia rufa, is composed of three polypeptides, designated Vn1 (175 kDa), Vn2 (160 kDa) and Vn3 (45 kDa) in SDS-polyacrylamide gel electrophoresis. Three subunits of vitellin were presented in the female adult hemolymph, ovary and egg extracts, but not observed in the male. This vitellin was purified from the eggs of P. rufa by the FPLC techniques, anion exchange chromatography and gel permeation chromatography. In nature, vitellin of P. rufa has molecular weight of 400 kDa. Western blot analysis using polyclonal antiserum against purified vitellin showed that the antiserum was reacted with the three polypeptides, Vnl, Vn2 and Vn3 from the female adult hemolymph, ovary and egg extracts. Amino acid residues at N-terminus of three subunits were sequenced. The N-terminal sequences of large subunits, Vnl and Vn2, were similar to each other, But, the N-terminal sequences of small subunits Vn3, did not have any signnificant homology with large subunits.

• Korean Journal of Veterinary Research
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• v.35 no.1
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• pp.87-94
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• 1995

Western immunoblot analysis of antigen of T sergenti merozoite revealed that the immunodominant proteins of this organism were characterized as the 18KD, 29KD, 34KD, 45KD and 105KD in Korea. The 34KD and 45KD among those immunodominant proteins of the parasite were isolated and their amino acid sequences from the $NH_2$-terminus were determined and synthesized. They respective polypeptides were cationized to enhance their antigenicity, fortified with Freund's adjuvant and tested for immunogenicity in rabbits and cattle. The results obtained were as follows; 1. Theileria sergenti merozoite antigen was shown in 120KD, 100KD, 66KD, 45KD, 34KD and 30KD in western immunoblot using serum of rabbits immunized with 34KD synthetic polypeptide and 70KD, 58KD, 55KD and 45KD using bovine serum. In western immunoblot, 45KD, 34KD and 30KD were recognized by immunized rabbits, and 50KD and 45KD by cattle sera immunized with 45KD synthetic polypeptide, respectively. 2. The ELISA utilizing the synthetic polypeptides demonstrated significant antibody response to the respective peptides. After the 2nd booster injection, an OD of 0.760(preimmunization 0.132) in rabbits and an OD of 0.645(preimmunization 0.488) to 34KD synthetic polypeptide in cattle were observed. In animals immunized with 45KD synthetic polypeptide, after the 2nd booster injection, an OD of 0.640(preimmunization 0.144) in rabbit, and an OD of 0.776 (preimmunization 0.477) in cattle were measured. 3. After the 2nd booster the reciprocal IFA titer was 1:64 in rabbits and 1:512 in cattle immunized with the 34KD synthetic polypeptide. The IFA titre was observed as 1:512 in rabbit and 1:1,024 in cattle in immunized with the 45KD synthetic polypeptide.

• Biotechnology and Bioprocess Engineering:BBE
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• v.6 no.2
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• pp.100-106
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• 2001

This investigation was performed to study the influence of benomyl on photosynthetic pigments and enzymes in soybean leaves. Chlorophyll and pheophytin levels were reduced by benomyl 45 days after greening. These results indicate that chlorophyll a and b, and pheophytin must be controlled by benomyl. SDS-PAGE analysis showed that 50 and 14.5 kD polypeptides represented as large and small subunits of rubisco. In the both of these subunits, the band intensity of the control was significantly higher than that after benomyl treatment, indicating that these two subunits are affected by benomyl. Benomyl strongly inhibited both the activity and content of rubisco as its concentration was gradually increased. However, it remains unclear whether this reduction of rubisco level was due to a reduced level of rubisco activase. Two major polypeptides of 46 and 42 kD were identified as rubisco activase subunits by SDS-PAGE. The intensity of these two bands was shown to be higher in the control than after benomyl treatment. These results indicate that the rubisco decrease resulting from increased benomyl concentrations was caused by rubisco activase. A significant decrease in both the activity and content of rubisco activase by benomyl was also observed. There results suggest that the decrease in rubisco level caused by benomyl is accompanied by a decrease in both the activity and content of rubisco activase.

• International Journal of Industrial Entomology and Biomaterials
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• v.15 no.1
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• pp.17-21
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• 2007

An indigenous multivoltine silkworm, Nistari was evaluated for their thermo tolerance by exposing the larvae to various temperature regimes for eight hours. Among different temperature exposed, this strain has significant tolerance at $32^{\circ}C$. Analysis of heat shock protein revealed the expression of 70 kDa and 64 kDa polypeptides in fat body and midgut tissues. Interestingly esterase isozyme pattern in midgut showed characteristic expression of Est-1 and Est-3 at different temperatures signifying role in heat and cold shock.

• Proceedings of the Korean Information Science Society Conference
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• 2012.06a
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• pp.397-399
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• 2012

생물학에서 특정 단백질을 구성하는 아미노산 서열을 결정하는 것은 매우 중요하다. 이를 위해 펩타이드의 단동위질량을 결정하는 것은 매우 중요한 문제이다. 하지만 Selenocysteine(Sec)이 가지고 있는 특이 성질 때문에 기존의 알고리즘으로 Sec를 포함한 펩타이드의 단동위질량을 결정하는 것은 쉽지 않다. J.W.Kim et.al.은 Sec의 특성을 고려하여 새로운 모델을 제시하고 그에 맞는 알고리즘을 보였다. 이 논문에서는 실험을 통해 J.W.Kim et.al.이 제안한 알고리즘을 실제 프로그램으로 구현하여 실행할 때 고려해야 할 주요 파라미터 및 함수에 대한 분석을 수행한다.

• Journal of Microbiology and Biotechnology
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• v.25 no.5
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• pp.658-661
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• 2015

Genes encoding enzymes with sequence similarity to hopanoids biosynthetic enzymes of other organisms were cloned from the hopanoid (hop) gene cluster of Streptomyces peucetius ATCC 27952 and transformed into Streptomyces venezuelae YJ028. The cloned fragments contained four genes, all transcribed in one direction. These genes encode polypeptides that resemble polyprenyl diphosphate synthase (hopD), squalene-phytoene synthases (hopAB), and squalene-hopene cyclase (hopE). These enzymes are sufficient for the formation of the pentacyclic triterpenoid lipid, hopene. The formation of hopene was verified by gas chromatography/mass spectrometry.

• Bulletin of the Korean Chemical Society
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• v.24 no.6
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• pp.757-762
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• 2003

Since theoretical investigations of gas phase clusters enable the evaluation of intrinsic molecular properties and intermolecular interactions, one can predict the macroscopic properties of bulk matter, from a microscopic determination of the properties of individual atoms, molecules, or clusters. Based on the insights obtained from theoretical investigations of the properties of a large number of cluster systems (ranging from simple water clusters to large π-systems), we have investigated the properties of various novel molecular systems including endo/exohedral fullerenes, nanotori, nonlinear optical materials, ionophores/receptors, polypeptides, enzymes, organic nanotubes, nanowires, and electronic and nano-mechanical molecular devices. The present minireview highlights some of the interesting results obtained in the course of our extensive theoretical investigations of clusters and nanomaterials.

• Journal of Microbiology and Biotechnology
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• v.4 no.2
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• pp.108-112
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• 1994

The genetic organization of the urease gene cluster from an alkalophilic Bacillus pasteurii was determined by subcloning and Tn5 transposon mutagenesis of a 10.7 kilobasepair cloned fragment. A region of DNA between 5.0 and 6.0 kb in length is necessary for urease activity. In vitro transcription-translation analysis of transposon insertion mutants of the cloned urease genes demonstrated that the major ($M_r$ 67,000) and minor ($M_r$ 20,000) structural peptides of urease are encoded at one end of the urease gene cluster and at least 3 additional polypeptides are encoded by adjacent DNA sequences.

• The Korean Journal of Zoology
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• v.22 no.3
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• pp.115-124
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• 1979

Two homotetrameric lactate dehydrogenase isozymes from Fluta alba and one from Ophicephalus argus were purified by combination of gel filtration and DEAE-cellulose chromatogrphy. The final preparations were isozymically pure and used to elicit antibodies in rabbits. The immunochemical reactivities demonstrated that the amino acids of active site is not to be included in the antigenic determinants, that antibodies or unknown component of immunized rabbit serum might be responsible for the electrophoretic abnormality and that two subunits share common antigenic determinants, reflecting that these polypeptides have a common evolutionary origin.