• Applied Biological Chemistry
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• 제30권2호
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• pp.163-168
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• 1987

전보에서 보고한 Bradyrhizobium japonicum 9 균주와 Rhizobium fredii 7 균주의 균체 중의 단백질 Pattern의 차이를 일차원 및 이차원 전기영동법에 의하여 조사하였다. 일차원 전기영동법 (SDS-PAGE)에 의해서는 두 group의 균체에서 모두 52개의 band가 관찰되었고 그 중 6개의 main band 로써 group 간의 차이가 확인되었으며, 이차원 전기영동법(2D-PAGE)에 의한 두 group간에 단백질 구성은 Rhizobium fredii 에서는 단백질이 산성 쪽에, Bradyrhizobium japonicum 는 alkali성 쪽에 비교적 많이 분포되어 있었다. 또한 두 group 간의 균체 아미노산 조성을 조사한 결과 조선상의 뚜렷한 차이가 없었다. 분리된 근류균을 확인하는데 전기영동법이 유용하였으며, 일차원 전기영동법은 많은 균주를 신속하게 확인할 수 있었고 이차원 전기영동법은 해상력 및 분리된 단백질 spot를 분석하는데 용이하였다.

• BMB Reports
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• 제32권5호
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• pp.445-450
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• 1999

We purified and characterized a bovine pregnancy-associated protein in pregnant cow urine using two-dimensional gel electrophoresis. Urine from cows was collected according to their status of pregnancy and non-pregnancy. Proteins in the cow urine were fractionated with 50% ammonium sulfate prior to two-dimensional gel electrophoresis. Proteins separated on the gels were compared in terms of expression level and new expression by molecular mass and isoelectric point. We localized two pregnancy-associated protein spots on the gels at molecular masses of 24 kDa and 20 kDa and isoelectric points of 5.5 and 5.7, respectively. Likewise, two non-pregnancy specific proteins were localized at 27 kDa and 28 kDa with isoelectric points of 5.7 and 5.9, respectively. To rule out the possibility that environmental or genetic factors might influence the expression of the proteins, we demonstrated the pregnancy-associated expression of the proteins in two-dimensional gels with pregnant urine taken from cows raised in a different institute. The pregnancy-associated protein with molecular mass of 20 kDa and isoelectric point of 5.7, namely spot 2, was microsequenced and found to be highly homologous to the bovine collagen alpha 1 chain.

• 대한의생명과학회지
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• 제23권4호
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• pp.395-401
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• 2017

Various preclinical and clinical trials have been conducted the efficacy of celastrol. In data presented in the current manuscript is the first trial to inhibit Helicobacter pylori with celastrol. In this study, the quantitative change of various H. pylori proteins including CagA and VacA by the anti-bacterial effect of celastrol was determined. The anti-H. pylori effects of celastrol was investigated by performing 2-dimensional electrophoresis and additional supporting experiments. After 2-dimensional electrophoresis analysis, spot intensities were analyzed and then each spot was identified using matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS) or peptide sequencing using Finnigan LCQ ion trap mass spectrometer (LC-MS/MS). The results show that celastrol has multiple effects on protein expression in H. pylori.

• Preventive Nutrition and Food Science
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• 제13권4호
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• pp.366-369
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• 2008

Proteins secreted from bifidobacteria are believed to play important roles in human intestines via interacting with different host cells. In this respect, proteins secreted from Bifidobacterium pseudocatanulatum BP1, which has been rarely studied, were analyzed by two-dimensional gel electrophoresis (2DE). Using this approach, approx-imately 21 protein spots on a 2DE gel were detected and 10 of these spots were identified by mass spectrometry. Five spots were identified as hypothetical proteins and the remaining 5 spots were identified as a putative iron-side-rophore binding lipoprotein, a short-chain dehydrogenase/reductase SDR, an exonuclease, cytochrome P450 hydroxylase, and a putative dehydrogenase. The identification of secreted putative iron-siderophore binding lipoprotein was highly interesting since it is an important protein that is involved in ferric iron uptake in pathogenic bacteria. This finding could accelerate studies on the probiotic effect of Bifidobacterium by explaining the competition between bifidobacteria and intestinal pathogens for ferric iron.

• Bulletin of the Korean Chemical Society
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• 제23권11호
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• pp.1511-1512
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• 2002

• Journal of Life Science
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• 제9권1호
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• pp.35-39
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• 1999

Elecrophoreticl analysis of a 36 kDa protein was runned by SDS-PAGE, isoelectric focusing (IEF) and two dimensional electrophoresis pattern. Major 36 kDa and 25, 46, 48, 66 kDa protein were detected by Coomassie blue stain on SDS-PAGE. Major 36kDa protein was eluted for production of antiserum for serological analysis, IEF and two dimensional electrophoresis. Isoelectric point of 36kDa was aout pH 8.5. Two dimensional electrophoresis of eluted 36kDa showed one point on the gel. Anti-36 kDa serum made by newzilland rabbit for serological test. In ELISA, final titer of antibody was 100×{TEX}$2^5}${/TEX} : 1. Neutralize ability of serum was examined by slide agglutination test and colonization test in rat. Anti-36 kDa serum agglutinated whole cell of V. vulnificus were inhibited colonization on intestine in rat. Accordingly In this paper contain some electrophoretical analysis and serological test of a 36 kDa OMP of V. vulnificus.

• Genomics & Informatics
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• 제9권4호
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• pp.197-199
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• 2011

The biological interpretation of two-dimensional (2D) gel electrophoresis experiments is a key step toward understanding the functions of biological systems. We here present a web-based integrated database, called 2DSpotDB, for the management of proteome data derived from several pathogens. The 2DSpotDB was established as a part of the management of a pathogen proteome project at the Korea National Institute of Health. The goals of the 2DSpotDB implementation are to store and define important pathogen genes, retrieve information obtained by 2D polyacrylamide gel electrophoresis and mass spectrometry, and create an integrated system to provide pathogen proteome information for biological scientists. This database currently contains 14 gels and information on 387 protein spots, among which 329 proteins were identified and annotated.

• 한국컴퓨터정보학회논문지
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• 제14권7호
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• pp.41-47
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• 2009

단백질 연구에 획기적인 기여를 한 2차원 전기영동법에 의한 결과영상을 처리하기 위하여 일반적으로 제안되었던 기존의 영상처리기법들이 가지는 문제점 중 하나인 단백질을 나타내는 여러 개의 스팟들이 겹치고 포화되고 있는 복잡영역에서 단백질 스팟을 하나로 인식하거나 검출이 불가능하여 오류를 범하게 되는 문제점에 대한 해법을 제안한다. 복잡영역내의 각 각점의 누적 기울기를 산출하고 누적 기울기 영상을 워터쉐드 기법으로 영역 분할하여 스팟을 분리한다. 그 결과 스팟 분리에 있어서 기존의 기법들을 적용하였을 때 보다 더 우수하고 효율적인 결과를 나타내며, 2차원 전기영동 결과영상에 숨겨져 있었던 단백질 스팟을 더 많이 검출할 수 있고, 예측의 범위 또한 확장시킨다.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.25-31
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• 2006

Changes of CP4 5-enolpyruvylshikimate-3-phosphate synthase (CP4 EPSPS) in the glyphosate-tolerant Roundup Ready soybean were examined using purified CP4 EPSPS produced in cloned Escherichia coli as a control. CP4 EPSPS in genetically modified soybean was detected by twodimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and electrospray ionization tandem mass spectrometry (ESI-MS/MS) with databases. CP4 EPSPS in soybean products was resolved on 2-DE by first isoelectric focusing (IEF) based on its characteristic pI of 5.1, followed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) based on its molecular mass of 47.5 kDa. We quantified various percentages of soybean CP4 EPSPS. The quantitative analysis was performed using a 2D software program on artificial gels with spots varying in Gaussian volumes. These results suggested that 2-DE image analysis could be used for quantitative detection of GM soybean, unlike Western blotting.

• Journal of Microbiology and Biotechnology
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• 제22권5호
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• pp.654-658
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• 2012

Osteoarthritis (OA) is the most common rheumatic pathology. One of the major objectives of OA research is the development of early diagnostic strategies such as those using proteomic technology. Synovial fluid (SF) in OA patients is a potential source of biomarkers for OA. The efficient and reliable preparation of SF proteomes is a critical step towards biomarker discovery. In this study, we have optimized a pretreatment method for twodimensional gel electrophoresis (2DE) separation of the SF proteome, by enriching low-abundance proteins and simultaneously removing hyaluronic acid, albumin, and IgG. SF samples pretreated using this optimized method were then evaluated by 1DE and 2DE separation followed by immunodetection of cartilage oligomeric matrix protein (COMP), a known OA biomarker, and by the identification of 3 proteins (apolipoprotein, haptoglobin precursor, and fibrinogen D fragment) that are related to joint diseases.