• 대한의용생체공학회:의공학회지
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• 제28권5호
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• pp.601-608
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• 2007

2 Dimensional Gel Electrophoresis(2DGE) is an essentialmethodology for analysis on the expression of various proteins. For example, information for the location, mass, expression, size and shape of the proteins obtained by 2DGE can be used for diagnosis, prognosis and biological progress by comparison of patients with the normal persons. Protein spot matching for this purpose is comparative analysis of protein expression pattern for the 2DGE images generated under different conditions. However, visual analysis of protein spots which are more than several hundreds included in a 2DGE image requires long time and heavy effort. Furthermore, geometrical distortion makes the spot matching for the same protein harder. In this paper, an iterative algorithm is introduced for more efficient spot matching. Proposed method is first performing global matching step, which reduces the geometrical difference between the landmarks and the spot to be matched. Thus, movement for a spot is defined by a weighted sum of the movement of the landmark spots. Weight for the summation is defined by the inverse of the distance from the spots to the landmarks. This movement is iteratively performed until the total sum of the difference between the corresponding landmarks is larger than a pre-selected value. Due to local distortion generally occurred in 2DGE images, there are many regions in whichmany spot pairs are miss-matched. In the second stage, the same spot matching algorithm is applied to such local regions with the additional landmarks for those regions. In other words, the same method is applied with the expanded landmark set to which additional landmarks are added. Our proposed algorithm for spot matching empirically proved reliable analysis of protein separation image by producing higher accuracy.

• 약학회지
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• 제50권4호
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• pp.268-271
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• 2006

Protein kinase UL97 is an unusual protein kinase that can phosphorylate nucleoside analogs as well as protein/peptide. Previously we found a H2B-derived peptide, KESYSVYVYKV and reported that the P+5 position (K) is important. To further understand the substrate specificity at the P+5 position, we introduced spot assay system and showed that a peptide containing K residue among other amino acids at the P+5 position is the best substrate. Also other residues such as M, I, L, or G are good enough to be substrate of UL97. This result may aid the discovery of a new antiviral inhibitor.

• 한국환경농학회지
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• 제32권1호
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• pp.48-54
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• 2013

수수 종자의 품종 간 특이적으로 발현하는 단백질을 동정하여 기능성 유전자를 확보하고 이들 유전자를 이용하여 수수의 기능성 강화 및 품종 판별 기술 개발을 위한 유용 유전자를 확보하고자 프로테오믹스 기법을 이용하여 수수 종자로부터 단백질을 추출하였다. 추출한 단백질을 이차원전기영동후, colloidal CBB 염색을 통해 품종 별로 발현에 차이를 보이는 단백질을 분석하였다. 총 652 개의 spot들 중에 8개의 단백질 spot들이 발현 정도에 변화를 보였으며, 이들 단백질을 MALDI-TOF/TOF MS와 MASCOT database를 통해 동정한 결과, RNA metabolism (spot 1, spot 4) HSP (spot 2), 저장 단백질 (spot 3, spot 5, spot 6), 산화-환원 (spot 8) 관련 단백질 등이 동정되었다. 특히 동정된 단백질은 주로 흰찰수수 (WCS)에서 발현 정도가 높게 나오는 경향을 보였으며, 흰찰수수 (WCS)에서 유일하게 발현 되는 단백질로 Cupin family protein, Gloubulin 등이 동정되었다. DEAD-box helicase는 흰찰수수 (WCS)를 제외한 나머지 세 품종에서 발현되었다. Ribonuclease T2와 Aldo-Keto reductase는 대풍수수 (DPS)를 제외한 나머지 세 품종에서 발현되었다. HSPs는 토종수수 (TJS)에서만 발현 되는 것을 확인하였다. 이들 동정된 단백질들은 수수의 품종 별 특성을 이해하는데 중요한 단서를 제공할 것으로 예측된다.

• International Journal of Industrial Entomology and Biomaterials
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• 제9권1호
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• pp.117-121
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• 2004

The patterns of diapause-associated proteins of silkworm eggs were analyzed by two-dimensional (2-D) gel electrophoresis. Among the hundreds of spots on the 2-D gels, at least two proteins were considered to be associated with diapause. A protein, spot 4, with an approximate molecular weight of 38 kDa and pI 6.1 was observed in the HCI-treated, cold-treated, and diapause eggs, respectively. Spot 4 was undetectable in unfertilized eggs and non-diapause eggs at two days after oviposition, suggesting that this protein may be associated with the entrance to diapause. A protein, spot 11, with an approximate molecular weight of 21 kDa and pI of 61 was detected in the unfertilized, HCl-treated, and cold-treated eggs, respectively, after oviposition by normal moths. In diapausing eggs, a protein corresponding to spot 11 was observed in 3-, 5-, and 30-day-old eggs, while the protein was not detected one-day-old eggs. The protein corresponding to spot 11 was not detected in unfertilized and non-diapause eggs obtained from subesophabeal ganglion (SG)-extirpated moths either. Spot 11 was also considered to be a diapause specific protein, which occurred at only early embryonic stage under the control of diapause-downregulated gene.

• BMB Reports
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• 제37권6호
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• pp.726-734
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• 2004

White spot disease (WSD) is caused by the white spot syndrome virus (WSSV), which results in devastating losses to the shrimp farming industry around the world. However, the mechanism of virus entry and spread into the shrimp cells is unknown. A binding assay in vitro demonstrated VP28-EGFP (envelope protein VP28 fused with enhanced green fluorescence protein) binding to shrimp cells. This provides direct evidence that VP28-EGFP can bind to shrimp cells at pH 6.0 within 0.5 h. However, the protein was observed to enter the cytoplasm 3 h post-adsorption. Meanwhile, the plaque inhibition test showed that the polyclonal antibody against VP28 (a major envelope protein of WSSV) could neutralize the WSSV and block an infection with the virus. The result of competition ELISA further confirmed that the envelope protein VP28 could compete with WSSV to bind to shrimp cells. Overall, VP28 of the WSSV can bind to shrimp cells as an attachment protein, and can help the virus enter the cytoplasm.

• Childhood Kidney Diseases
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• 제4권1호
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• pp.1-5
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• 2000

목 적 : 24시간 소변내 단백량을 예측하는데 있어 spot urine의 단백은 소변량에 따른 보정이 필요하여 spot urine의 protein/creatinine(P/Cr)비를 이용하여왔다. 하지만 크레아티닌 측정에 많은 시간과 비용이 소모되는 단점이 있고, 비중(specific gravity; SG)을 이용하여 creatinine(estimated creatinine; Cr-est)을 산출할 수 있다는 보고가 있어, 24시간 소변내 단백량을 예측하는데 P/Cr-est비의 유용성을 알아보고자 본 연구를 시행하였다. 방 법 : 1998년 3월부터 6월까지 고려대학교 의료원 구로병원 소아과에 내원한 환아 147명을 대상으로 spot urine에서 단백, creatinine과 비중을 측정하여 creatinine과 비중의 상관관계 및 그 관계식을 구하고 24시간 소변내 단백량과 비교하였다. 결 과 : 1) spot urine의 creatinine과 비중은 통계적으로 의의 있는 상관관계를 보였다(R=0.407, P<0.001, Cr=SG. x 4485.82-4482.87). 2) 24시간 소변내 단백량은 P/Cr비와 통계적으로 의의 있는 상관관계를 보였다(R=0.771, P<0.001, 24시간뇨중 단백=0.338 x (P/Cr) +667.885). 3) 24시간 소변내 단백 량은 P/Cr-est 비와 통계적으로 의의 있는 상관관계를 보였다. (R=0.723, P<0.001, 24시간뇨중 단백=0.354 x (P/Cr-est) +726.044) 결 론 : 소아단백뇨를 예측하는데 있어 spot urine의 비중으로 산출한 creatinine치를 이용하여 얻어진 P/Cr-est비가 선별검사로서 유용하게 사용될 수 있으리라 생각된다.

• 한국임상수의학회지
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• 제36권1호
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• pp.85-87
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• 2019

Eight-week-old Spot-billed duck were presented with visible drooping of both of wings. On physical examination, the Spot-billed ducks revealed valgus deformity of the carpal joint resulting in the primary flight feathers protruding dorsally. The bird was in good body condition and there was no loss of motion in any of the joints in the wings. The bird was fed chicken pellet with 18.5% of protein level and reared in a cage. Based on the clinical presentation and physical examination 'angel wing' was diagnosed. Wing bandage and nutritional change to lower-protein diet with fresh vegetables were applied simultaneously. And duck was transferred to wider outside pen with small pond. Four-week afterward clinical signs of angel wing were improved.

• 한국약용작물학회지
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• 제22권5호
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• pp.398-404
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• 2014

Salt stress is one of the major abiotic stresses affecting the yield of ginseng (Panax ginseng C. A. Meyer). The objective of this study was to identify bio-marker, which is early responsive in salt stress in ginseng, using proteomics approach. Ginseng plants were exposed to 5 ds/m salt concentration and samples were harvested at 0, 6, 12 and 18 hours after exposure. Total proteins were extracted from ginseng leaves treated with salt stress using Mg/NP-40 buffer and were separated on high resolution 2-DE. Approximately $1003{\pm}240$ (0 h), $992{\pm}166$ (6 h), $1051{\pm}51$ (12 h) and $990{\pm}160$ (18 h) spots were detected in colloidal CBB stained 2D maps. Among these, 8 spots were differentially expressed and were identified by using MALDI-TOF/TOF MS or/and LC-MS/MS. Ethylene response sensor-1 (spot GL 1), nucleotide binding protein (spot GL 2), carbonic anhydrase-1 (spot GL 3), thylakoid lumenal 17.9 kDa protein (spot GL 4) and Chlorophyll a/b binding protein (spot GL 5, GL 6) were up-regulated at the 12 and 18 hour, while RuBisCO activase B (spot GL 7) and DNA helicase (spot GL 8) were down-regulated. Thus, we suggest that these proteins might participate in the early response to salt stress in ginseng leaves.

• 한국정보과학회논문지:컴퓨팅의 실제 및 레터
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• 제16권5호
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• pp.551-555
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• 2010

단백질 2-DE 이미지 분석에서 단백질 자체가 가지고 있는 불안정성과 2-DE 실험이 가지고 있는 근본적인 문제점으로 인하여 이미지 스팟 매칭 분석의 정확도가 낮아지게 된다. 이 논문에서는 다중 참조이미지를 사용하여, 스팟 매칭 패턴의 정확도에 큰 영향을 주는 이미지 찌그러짐을 보완하고, 그에 따른 노이즈 스팟 제거와 참조 이미지 품질에 의한 정확도 저하를 최소화하는 방법을 제안하였다. 또한 2-DE 이미지의 데이터 특성에 의하여 이미지 수가 증가할 때 성능이 급격히 떨어지는 문제를 해결하기 위하여, 다중 참조이미지를 사용하여 구축한 스팟 매칭 데이터베이스를 이미지의 생물학적 특성에 의하여 "분할 및 확장" 방법을 사용하여, 정확도를 향상시키는 동시에 패턴 길이를 보장하는 스팟 매칭 패턴을 효과적으로 생성하였다. 실험에서는 실제 인간 2-DE 이미지 데이터를 사용하여 제안한 방법의 타당성을 보여준다.

• 한국식품과학회지
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• 제43권1호
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• pp.12-16
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• 2011

본 연구에서는 프로테오믹스 기술을 활용하여 칼슘함량이 증가된 새송이버섯과 일반 새송이 버섯에서 단백질 발현의 변화를 조사하였다. 또한 현저한 차이를 보이는 단백질들을 분리, 동정함으로써 새송이버섯 칼슘강화의 기작규명에 기초자료를 제공하고자 하였다. 새송이버섯의 단백질 패턴을 확인한 결과 15 Kda에서 100 Kda 사이에 존재하는 60% 정도의 spot은 산성 pI를 가지며 나머지 40% 정도의 spot은 염기성 영역에 나타났다. 100 Kda 이상에서는 polypeptide spot이 거의 나타나지 않았다. 그리고 9.0 이상의 pI에서도 spot은 거의 나타나지 않았다. 두 배 이상의 발현변화를 보이는 30여개의 spot들 중 10개의 spot에 대한 단백질동정을 할 수 있었다. 10개의 확인된 단백질은 8개의 단백질은 발현이 증가하는 것으로 나타났으며 2개의 단백질은 발현이 감소하는 것으로 나타났다. 동정된 단백질들의 기능을 고찰한 결과 새송이버섯의 칼슘강화기작을 규명하는데 기초자료로 활용할 수 있을 것으로 기대된다.