• Journal of Microbiology
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• 제37권1호
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• pp.21-26
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• 1999

Two novel calcium-binding proteins, named CAB-I and CAB-II, have been isolated from Streptomyces coelicolor. Purification of the calcium-binding proteins involved heat treatment, fractionation with ammonium sulfate, acid treatment, anion exchange and hydrophobic interaction column chromatography, FPLC gel filtration, and preparative isoelectric focusing. A chelex competitive assay and 45Ca autoradiography verified the calcium-binding ability of the proteins. The major band CAB-II has an apparent molecular weight of 26,000 determined by SDS-polyacrylamide gel electrophoresis and 340,000 determined by gel filtration. The isoelectric point of this molecule showed the acidic nature of the molecule. N-terminal amino acid sequence analysis shows homology to rat Ca2+/calmodulin-dependent protein kinase-II (CAB-II) and yeast phosphoprotein phosphatase (CAB-I).

• The Plant Pathology Journal
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• 제16권1호
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• pp.9-18
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• 2000

Using differential display techniques, a new acidic pathogenesis-related (PR) protein-1 cDNA (GMPRla) gene was isolated from a cDNA library of soybean (Glycinemax L.Merr, cultivar Jangyup) hypocotyls infected by Phytophthora sojae f. sp. glycines. The 741 bp of fulllength GMPRla clone contains an open reading frame of 525 nucleotides encoding 174 amino acid residues (pI 4.23) with a putative signal peptide of 27 amino acids in the N-terminus. Predicted molecular weight of the protein is 18,767 Da. The deduced amino acid sequence of GMPRla has a high level of identity with PR-1 proteins from Brassica napus, Nicotiana tabacum, and Sambucus nigra. The GMPRla mRNA was more strongly expressed in the incompatible than the compatible interaction. The transcript accumulation was induced in the soybbean hypocotyls by treatment with ethephon or DL-$\beta$-amino-n-butyric acid, but not by wounding. In situ hybridization data showed that GMPRIa mRNAs were usually localized in the vascular bundle of hypocotyl tissues, especially phloem tissue. Differences between compatible and incompatible interactions in the timing of GMPRla mRNA accumulation were remarkable, but the spatial distribution of GMPRla mRNA was similar in both interactions. However, more GMPRla mRNA was accumulated in soybean hypocotyls at 6 and 24 h after inoculation.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.303-310
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• 1999

A cost-effective mass production method for a strong antimicrobial peptide, buforin II, which was isolated from the stomach of Bufo bufo gargarizans, has been developed. This method is based on the neutralization of the positive charge of buforin II by fusion with a cysteine-rich acidic peptide (CAP) to avoid any lethal effect on the host. The neutralized fusion peptide was multimerized and expressed in Escherichia coli as tandem repeats to increase the production yield. Multimers of the CAP-buforin II fusion peptide were successfully expressed at high levels in E. coli as inclusion bodies. More than 100mg of pure buforin II was obtained per 11 of E. coli culture after cleaving the multimeric polypeptide with CNBr. The buforin II obtained from the recombinant E. coli had antimicrobial activity identical to that of natural buforin II. The proposed expression system can provide a cost-effective mass production method for both antimicrobial peptides and other host-lethal basic proteins.

• Archives of Pharmacal Research
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• 제25권5호
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• pp.704-708
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• 2002

A fast and sensitive protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, Coomassie Brilliant Blue R-250 (CBBR) and a basic dye, Neutral Red (NR) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution enhances the staining effect of CBBR on protein bands, and also reduces gel background. It is a rapid staining procedure, involving fixing and staining steps with short destaining that are completed in about 1 h. As the result, it showed two to fourfold increase in sensitivity comparing with CBBR staining. The stained protein bands can be visualized at the same time of staining.

• 자연과학논문집
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• 제10권1호
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• pp.27-32
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• 1998

HIV-1 gp41의 세포내 부분과 상호작용하는 단백질 유전자를 분리할 목적으로 yeast two hybrid system을 사용하여 검색하였다. 전체 $1.4 \times 10^6 colony를 검색하여 최종적으로 20개의 colony를 얻었다. 이들 colony로부터 분리된 유전자의 염기배열을 결정하여 본 결과, acidic ribosomal protein P0, beta tubulin, alpha catenin등의 세가지 종류임을 밝혔다. 이들은 yeast system 내에서 매우 특이적으로 gp41과 상호작용하고 있음을 알았다.

• Bulletin of the Korean Chemical Society
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• 제18권1호
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• pp.18-23
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• 1997

Data from structure/activity studies in vir gene induction system have led to evaluate the working hypothesis of interaction between phenolic inducers and phenol binding proteins. The primary specificity in the association of a phenolic inducer with its receptor in our system is hypothesized to be the hydrogen bonding interactions through the ortho methoxy substituents as well as the proton transfer between the inducer and the binding protein. In this paper the proposed working model for phenol-mediating signal transduction was evaluated in several ways. The importance of the general acid-base catalysis was first addressed by the presence of an acidic residue and a basic residue in the phenol binding protein. Series of compounds were tested for vir gene expression activity to confirm the generation of a strong nucleophile by an acidic residue and an involvement of a basic residue as a proton acceptor. An attempt was made to correlate the pKa values of the phenolic compounds with vir gene induction activities as inducers to further support the proposed proton transfer mechanism. Finally, it was also observed that the regioselectively attached methoxy group on phenol compounds is required as the proper hydrogen bond acceptor.

• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.313.2-313.2
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• 2002

Environmentally benign protein staining method in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) using both an acidic dye, zincon (ZC) and a basic dye. ethyl violet (EV) is described. It is based on a counter ion-dye staining technique that employs oppositely charged two dyes to form an ion-pair complex. The selective binding of the free dye molecules to proteins in an acidic solution produces bluish violet colored bands. It is a rapid procedure, involving only fixin and staining steps that are completed in 45 min. (omitted)

• 대한치과교정학회지
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• 제32권6호
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• pp.443-453
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• 2002

본 연구의 목적은 다양한 교정용 브라켓의 표면에 형성되는 타액성 피막의 조성을 확인하고, 전타액, 악하선타액 및 이 하선타액에서 유래하는 타액성 피막의 성분을 비교하는 것이다. 네 가지 서로 다른 종류의 교정용 브라켓을 본 연구에 사용하였다. 이들은 $022{\times}028$ Roth Prescription의 상악 소구치 브라켓으로 조성은 다음과 같다: 스테인레스 스틸, 단결정 사파이어, 다결정 알루미나 및 플라스틱 브라켓. 교정용 브라켓을 각각 전타액, 이하선타액 및 악하선타액에 2시간 배양하여 타액성 피막을 형성시켰다 브라켓 피막의 타액성분은 sodium dodecyl sulfate-polyacrylamide gel electrophoresis, Western transfer method 및 면역검출법을 통해 확인하였다. 이 결과 low-molecular weight salivary mucin, ${\alpha}-amylase$, secretory IgA (sIgA), acidic proline-rich proteins, cystatins 등이 모든 브라켓의 타액성 피막에 존재하였으며, 치아우식증의 원인균인 Streptococcus mutans의 부착을 촉진시키는 타액단백질인 high-molecular weight mucin은 어떤 브라켓에도 부착하지 않았다. 그러나, 비록 동일한 타액단백질이 모든 브라켓에서 발견되었지만, 타액단백질 부착 양상은 타액의 종류 및 브라켓의 종류에 따라 양적 및 질적으로 다르게 나타났다. 특히 sIgA는 이하선타액에서 유래한 브라켓 피막에 더 많이 부착하였고, cystatins의 경우는 플라스틱 브라켓에서 유래한 브라켓 피막에 더 많이 존재하였다 본 연구는 다양한 타액단백질이 교정용 브라켓에 부착하며, 타액단백질이 타액의 출처 및 브라켓의 종류에 따라 교정용 브라켓의 표면에 선택적으로 부착함을 나타내었다.

• 한국식품과학회지
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• 제30권4호
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• pp.988-991
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• 1998

소와 돼지 혈액중의 혈장 단백질을 분리하여, 성분 분석과 단백질 함량을 정량했으며 혈장단백질의 식품 기능성에 관하여서는 기존 유화제로서 활용되는 유청 단백질인 WPC와 비교하여 유화능력을 비교 검토하였다. FPLC 및 SDS-PAGE에 의한 성분분석 결과 혈장 알부민(SA)은 돈혈장단백질(PPP), 우혈장단백질(BPP), 유장단백질(WPC)의 순서로 많이 함유되었으며, 혈장 단백질에는 ${\beta}-globulin$분획(주로 transferrin)이 다량 포함되어 있고, 그외 미량성분(fibrinogen, immunoglobulin)들이 확인되었다. 단백질 함량은 BPP (85%) 및 PPP (82%) 모두 높은 함량을 보여 우수한 단백질원으로 이용 가능할 것으로 보였다. 유화력도 혈장단백질들이 단백질 농도 2% 이하에서는 WPC보다 높았으며, 4% 이상에서는 WPC보다는 약간 낮았다. 또한 염농도 및 pH 의존성에 관하여 유화력에 미치는 영향을 검토한 결과, PPP 및 BPP의 pH 영역에 대한 유화활성은 WPC와는 상이하게 산성쪽의 pH에서 염기성쪽보다 더 높은 활성을 보였으며, 염(NaCl)첨가로 인한 유화 활성의 영향은 WPC와 비교하여 pH 의존성이 상당히 높았으며, 특히 산성쪽에서 높은 활성을 나타냈다. 이상의 결과들을 살펴볼 때 소 및 돼지의 혈액에서 제조한 혈장단백질들은 우수한 유화특성을 갖는 식품소재로 확인되었다.

• BMB Reports
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• 제40권2호
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• pp.212-217
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• 2007

The chicken oviduct is a dynamic organ that produces secretory proteins such as ovalbumin during the laying period. In this study, we identified oviduct-specific proteins in hens during the egg-laying period by proteomic analysis. Proteins extracted from the magnum of hens of different ages (5, 35, and 65 weeks) were analyzed by two-dimensional gel electrophoresis to compare the intensity of proteins among samples. Approximately 300 spots were detected on each gel. Based on the comparison of image gels, we found that the intensity of eight spots in 35-week magnums was increased at least by 2-fold compared with the others. Five of the eight spots were identified as calumenin, acidic ribosomal phosphoproteins (ARP), prohibitin, heart fatty acid-binding protein, and anterior gradient-2 (AGR-2). In particular, ARP and AGR-2 were highly expressed in 35- week magnums compared with 5- and 65-week magnums. In addition, the level of these proteins was consistent with their RNA levels. Expression of AGR-2 mRNA was detected in the mature magnum, whereas no signal was observed in premature tissue. Among various tissues, expression of AGR-2 mRNA was highest in the magnum, high in the isthmus, and five fold lower in muscle. It was undetectable in the liver and in other tissues (heart and kidney). However, the mRNA levels of other proteins were ubiquitous among tissues. In transcriptional activity of AGR-2, a 3.0 kb fragment of promoter region containing potential estrogen receptor binding sites had enhanced its activity strongly. In conclusion, these results suggest that AGR-2 has functional regulatory roles in the chicken oviduct during the egglaying period.