• BMB Reports
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• v.37 no.4
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• pp.466-473
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• 2004

A new acid deoxyribonuclease (DNase) was purified from the cultured mycelia of Cordyceps sinensis, and designated CSDNase. CSDNase was purified by $(NH_4)_2SO_4$ precipitation, Sephacryl S-100 HR gel filtration, weak anion-exchange HPLC, and gel filtration HPLC. The protein was single-chained, with an apparent molecular mass of ca. 34 kDa, as revealed by SDS-PAGE, and an isoelectric point of 7.05, as estimated by isoelectric focusing. CSDNase acted on both double-stranded (ds) and single- stranded (ss) DNA, but preferentially on dsDNA. The optimum pH of CSDNase was pH 5.5 and its optimum temperature 55. The activity of CSDNase was not dependent on divalent cations, but its enzymic activity was inhibited by high concentration of the cation: $MgCl_2$ above 150 mM, $MnCl_2$ above 200 mM, $ZnCl_2$ above 150 mM, $CaCl_2$ above 200 mM, NaCl above 300 mM, and KCl above 300 mM. CSDNase was found to hydrolyze DNA, and to generate 3-phosphate and 5-OH termini. These results indicate that the nucleolytic properties of CSDNase are essentially the same as those of other well-characterized acid DNases, and that CSDNase is a member of the acid DNase family. To our knowledge, this is the first report of an acid DNase in a fungus.

• Proceedings of the Korean Society of Sericultural Science Conference
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• 2003.10a
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• pp.95-96
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• 2003

We investigated the proteins that are associated with initiation, maintenance, and termination of diapause in silkworm eggs by means of 2D-gel electrophoresis. 1. Materials : Bombyx mori: unfertilized eggs, diapause eggs, non-diapause eggs, cold-treated egges. 2. Methods: isoelectric focusing, SDS-PAGE, silver staining. (omitted)

• The Korean Journal of Zoology
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• v.36 no.2
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• pp.193-199
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• 1993

The protein po;vmorphisms and allele frequencies of wild house rat (Rattus norvegicus) population in Korea were studied. The studied proteins and enzymes were transferrin (Tf), albumin (Alb), fumarate hvdratase (FH), phospho!loucomutase (PGM), lactate dehvdrogenase A (LDHA) and lactate dehvdrogenase B (LDHB). There were two transferrin alleles, TP and Tf in Korean wild house rat popu1ation. The Tf2 allele was found for the first time by a starch gel, and confirmed by a polvacrvlamide gel isoelectric focusing and immunoblotting. The allele frequencies of TP and TF were 0.985 and 0.015, respectively. Two common alleles fumarate hydratase, FHa and FHb were found, and frequencies of FHa and FPP were calculated to be 0.714 and 0.286, respectively. The kequenw of FH in Korean wild house rat was higher than that of Finnish and Czechoslovakian population. Alb, PGM, LDHA and LDHB are only one phenotype each and all. Therefore, these proteins seem to be monomorphic in Korean wild house rat population.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.60 no.1
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• pp.29-32
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• 2015

Two-dimensional electrophoresis (2-DE) was executed to separate the seed storage proteins from the buckwheat. The proteins extracted from the whole seed proteins were better separated and observed in the use of lysis buffer. Using this method, the highly reproducible isoelectric focusing (IEF) can be obtained from polyacrylamide gels, and IEF from the polyacrylamide gel at all the possible pH range (5.0-8.0) was more easily separated than IPG (immobilized pH gradient) gels. The polyacrylamide gels in the first dimension in 2-DE was used to separate and identify a number of whole seed proteins in the proteome analysis. In this new apparatus using 2-DE, 27cm in length of plate coated with polyacrylamide gel was used and the experiment was further investigated under the various conditions.

• Microbiology and Biotechnology Letters
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• v.22 no.6
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• pp.627-635
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• 1994

The $\beta$-xylosidase from Penicillium sp. FX-102 was purified by 40~80% ammonium sulfate saturation, CM-Cellulose column chromatography, Sephadex G-200 gel filtration, and isoelec- tric focusing. The optimum pH and temperature for the activity of the $\beta$-xylosidase was pH 4.5 and 50$\circ$C, respectively. The enzyme was stable at the pH range of 4.5~5.5, and at 55$\circ$C for 10 min. The molecular weight of the enzyme was estimated to be about 300,000 daltons by Sephadex G-200 gel filtration and 310,000 daltons of monomer by SDS polyacrylamide gel electrophoresis. Isoelectric point of the enzyme was determined to be pH 4.4. The enzyme activity was strongly inhibited by Hg$^{2+}$, Ag$^{2+}$, n-bromosuccinimide and p-chloromercuribenzoate. Xylobiose (10 mM) was completely decomposed to xylose after 8 hrs enzyme reaction with 2 units of the $\beta$-xylosidase.

• Journal of Periodontal and Implant Science
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• v.35 no.2
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• pp.271-275
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• 2005

본 연구는 치주낭에 biofilm형태로 부착되어 질환을 유발시키고 항생제 빚 항균제에 저항을 일으키는 세균 독성요소를 규명하기 위해 시행된 기초연구이다. 치주질환의 주 병원균의 하나인 Porphyromonas gingivalis 381 biofilm의 세포외막에 특이하게 발현되는 단백질을 규명하기 위한 기초적인 자료를 얻기 위해 시행하였다. Porphyromonas gingivalis 381을 통상적인 세균 배양용 broth를 사용하여 혐기성 세균 배양기로 24시간 배양한 것을 대조군으로 하고, tissue culture plate를 이용하여 혐기성 배양조건 하에서24시간동안 biofilm을 형성하여 실험군으로 설정하였다. 세균을 수획하여 세포외막을 분리하고 isotonic isoelectric focusing을 시행한 결과 주로 약 20-30 kilodaltons에 해당하는 수종의 세균세포막 단백질이biofilm으로 배양한 세균에서 더 상승적으로 발현됨이 관찰되었고, 상이한 수종의 단백질도 planktonic culture broth로 배양한 세균에서 다 상승적으로 발현됨을 관찰할 수 있었다. 이것은 세균의 배양조건과 환경에 따라 그 외막 단백질이 서로 다르게 발현됨을 입증하는 기초적인 자료로서 향후 단백질의 동정과 성격을 규명하는 근간 실험으로 추진할 계획이다.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.230.3-231
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• 2003

The therapeutic use of protein pharmaceuticals produced by recombinant DNA technology is increasing in recent decades. In order to investigate the quality of recombinant proteins, it is important to identify and assign the impurities produced in the process of recombination or in storage conditions. Capillary Electrophoresis is emerging technology exhibiting high sensitivity, selectivity and speed and may be most powerful tools for this application. In this study, human growth hormone (hGH) has been analyzed by various mode of capillary electrophoresis such as capillary zone electrophoresis (CZE), capillary gel electrophoresis (CGE), and capillary isoelectric focusing (cIEF) to indicate the chemically or biologically oriented variants and the degraded fragments. (omitted)

• Applied Biological Chemistry
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• v.22 no.1
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• pp.1-9
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• 1979

The physical and chemical properties of ${\gamma}-conglycinin$ of soybean (Glycine max) were investigated. The soybean protein extracted from soybean meal using 0.2M NaCl solution at pH 4.5 was passed through a Sephadex G-150 column to isolate 7S globulin. ${\gamma}$-Conglycinin was isolated and purified from the 7S globulin with a DEAE Sephadex A-50 column chromatography. The protein preparation was pure on immunoelectrophoresis, polyacrylamide gel electrophoresis and gel isoelectric fouling. It had an isoelectric point at pH 5.4 and contained 16.12% nitrogen, 4.18% mannose and 1.21% glucosamine. Amino acid composition, in general, shaved that ${\gamma}-conglycinin$ contained higher contents of lysine, dicarboxylic acids and ammonia nitrogen, and lower contents of sulfur-containing amino acids and tryptophan. The subunits of ${\gamma}-conglycinin$ were distributed in the range of pH 4.6-5.5. The subunits located in the pH region of 4.6-5.0 and 5.0-5.5 were glycopeptides (molecular weight of 38,000) and simple peptide (MW of 32,000), respectively.

• Journal of Life Science
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• v.24 no.2
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• pp.186-190
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• 2014

Carbonic anhydrase isozymes (CAs) are widespread zinc-containing metalloenzyme family. The enzyme catalyzes the reversible interconversion of $CO_2$ and $HCO_3$. This reaction is the main role of CA enzymes in physiological conditions. CA III, one of the CA isozymes, has been identified in many tissues. It is distinguished from the other isozymes by several characteristics, particularly by a lower specific activity and by its resistance to acetazolamide. However, the physiological function of CA III in fish is unknown. In this study, we examined the detection of CAs in the Shaggy sea raven Hemitripterus villosus, using SDS-PAGE, isoelectric focusing (IEF), and western blot analysis. We detected a significant protein band with molecular weight about 30 kDa from the tissues of H. villosus by SDS-PAGE and western blotting. A specific band of CA III with pI 7.0 was detected by IEF and western blotting in gill and muscle. The immunoreaction of anti-CA III expressed in the gill of H. villosus was much stronger than other tissues. One explanation for this result is that the fish gill is the only organ that is exposed to the external environment and that plays an important role in acid-base relevant ion transfer, the transfer of $H^+$ and/or $HCO{_3}^-$, for the maintenance of systemic pH. This is the first report on the identification of a carbonic anhydrase III-like protein from H. villosus.

• The Korean Journal of Pharmacology
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• v.15 no.1_2 s.25
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• pp.1-5
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• 1979

Previous studies concerning the usefulness of pleural fluid glucose levels in differentiating causes of pleural effusions have been conflicting. Gelenger and Wiggers (1949), Calnan et al(1951) and Barber et al(1957) concluded that the lower the level of pleural fluid glucose, the more likely was tuberculosis, and that tuberculosis was unlikely if the pleural fluid glucose level was more than 80 mg/100 ml. Light and Ball(1973), however, reported that in the great majority of tuberculous pleural fluids the glucose concentration was high rather than low, concluded that the pleural fluid glucose levels were not useful in the differential diagnosis of pleural effusion. In this study, pleural fluid glucose was determined in 46 pleural effusions from various causes to evaluate the usefulness in the differential diagnosis of pleural effusion. In addition, the protein concentration and the electrophoretic patterns of protein and amylases in pleural fluid was compared with that of serum. And the results were as follows. 1. The mean glucose concentration of pleural fluid was 80.8 mg/100 ml in 22 tuberculous origin, 92.5 mg/100 ml in 12 cancer patient and 70.4 mg/100 ml in 10 undiagnosed cases. In 2 cases of paragonimiasis the pleural fliud glucose levels were low (mean, 32.0 mg/100 ml). The percentage of pleural fluid protein to serum is about 75% in all disease groups and the protein level of tuberculous pleural fluid was significantly correlated with that of serum. 2. The disc eletrophoretic patterns of pleural fluid were almost similar with that of serum in all disease groups but the prealbumin fraction was not observed in pleural fluid. 3. With the isoelectric focusing, 4 to 7 isoamylase was observed in serum and the isoelectric point was ranged from pH 5.8 to 7.8 and isoelectic point of main fracticn is pH 7.2. The isoelectic focusing patterns of amylase of pleural fluid were identical to that of serum in all disease group. With the above results it is concluded that the pleural fluid is exudate of serum and that the glucose levels of pleural fluid are not useful in the differential diagnosis of pieural effusions.