• 한국생물공학회:학술대회논문집
• /
• 한국생물공학회 2005년도 생물공학의 동향(XVI)
• /
• pp.47-47
• /
• 2005

There are many different surface technologies currently applied for preparation of protein chips. However, it requires innovative surface chemistry for capture proteins to be immobilized on chip surface keeping their conformation and activity intact and their orientation right, while they bind tightly and densely in a given array spot. Proteogen has developed 'ProteoChip BP' coated with novel proprietary linker molecules $(ProLinker^{TM})$ for efficient and robust immobilizations of capture proteins by improving surface properties of molecular captures. It was demonstrated that $ProLinker^{TM}$ gave the best surface performance in preparation of protein microarray chip base plates among others currently available on the market. In particular, the $ProLinker^{TM}-based$ surface chemistry has demonstrated to provide excellent performance in preparation of 'Antibody Chip' for analysis of biomarkers as well as proteome expression profiles. The linker molecule has also shown to be well applicable for development of biosensors and micro-beads as well as protein microarray and nano-array. ProteoChip BP can be used either for preparation of high-density array by using a microarrayer or for preparation of 'Well-on-a-Chip' with low density array, which is better applicable for quantitative analysis of biomarkers or protein-protein interactions. The biomarker assay can be performed either by direct or sandwich methods of fluorescence immunoassay. Application of ProteoChip BP has been well demonstrated by the extensive studies of 1) tumor-marker assays, 2) new drug screening by using 'Integrin Chip' and 3) protein expression profile analysis. Some of experimental results will be presented.

• 한국초지조사료학회지
• /
• 제34권4호
• /
• pp.262-268
• /
• 2014

In order to reveal the aluminum (Al) stress tolerance mechanisms in alfalfa plant at low pH soil, a proteomic approach has been conducted. Alfalfa plants were exposed to Al stress for 5 days. The plant growth and total chlorophyll content are greatly affected by Al stress. The malondialdehyde (MDA) and $H_2O_2$ contents were increased in a low amount but free proline and soluble sugar contents, and the DPPH-radical scavenging activity were highly increased. These results indicate that antioxidant activity (DPPH activity) and osmoprotectants (proline and sugar) may involve in ROS ($H_2O_2$) homeostasis under Al stress. In proteomic analysis, over 500 protein spots were detected by 2-dimentional gel electrophoresis analysis. Total 17 Al stress-induced proteins were identified, of which 8 protein spots were up-regulated and 9 were down-regulated. The differential expression patterns of protein spots were selected and analyzed by the peptide mass fingerprinting (PMF) using MALDI-TOF MS analysis. Three protein spots corresponding to Rubisco were significantly down-regulated whereas peroxiredoxin and glutamine synthetase were up-regulated in response to Al stress. The different regulation patterns of identified proteins were involved in energy metabolism and antioxidant / ROS detoxification during Al stress in alfalfa. Taken together, these results provide new insight to understand the molecular mechanisms of alfalfa plant in terms of Al stress tolerance.

• Molecules and Cells
• /
• 제40권9호
• /
• pp.685-695
• /
• 2017

Obesity and the metabolic disorders that constitute metabolic syndrome are a primary cause of morbidity and mortality in the world. Nonetheless, the changes in the proteins and the underlying molecular pathways involved in the relevant pathogenesis are poorly understood. In this study a proteomic analysis of the visceral adipose tissue isolated from metabolically healthy and unhealthy obese patients was used to identify presence of altered pathway(s) leading to metabolic dysfunction. Samples were obtained from 18 obese patients undergoing bariatric surgery and were subdivided into two groups based on the presence or absence of comorbidities as defined by the International Diabetes Federation. Two dimensional difference in-gel electrophoresis coupled with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry was carried out. A total of 28 proteins were identified with a statistically significant difference in abundance and a 1.5-fold change (ANOVA, $p{\leq}0.05$) between the groups. 11 proteins showed increased abundance while 17 proteins were decreased in the metabolically unhealthy obese compared to the healthy obese. The differentially expressed proteins belonged broadly to three functional categories: (i) protein and lipid metabolism (ii) cytoskeleton and (iii) regulation of other metabolic processes. Network analysis by Ingenuity pathway analysis identified the $NF{\kappa}B$, IRK/MAPK and PKC as the nodes with the highest connections within the connectivity map. The top network pathway identified in our protein data set related to cellular movement, hematological system development and function, and immune cell trafficking. The VAT proteome between the two groups differed substantially between the groups which could potentially be the reason for metabolic dysfunction.

• 대한수의학회지
• /
• 제45권2호
• /
• pp.155-161
• /
• 2005

Proteomics is a useful approach to know protein expression, post-translational modification and protein function. We investigated the protein expression pattern and identity in chickens fed with the tissue culture medium waste after harvest of Korean wild ginseng (TCM-KWG) (Panax ginseng). Two groups (n=60/group) of day old broiler chickens were administered with 0 (control) and 0.8% (treatment) TCM-KWG through drinking water. After 5 weeks, we examined the protein expression pattern of fibularis longus and superficial pectoral muscle by Two-dimensional electrophoresis analysis. Interestingly, TCM-KWG treatment significantly increased five spot's density, and markedly reduced five spot's density in the muscles. We identified 10 proteins (desmin, myosin light chain 1, heat shock 25 kDa protein, collapsin response mediator protein-2A, alpha enolase, vimentin, actin alpha 1, my023 protein, pyruvate kinase and troponin T) by the matrix-assisted laser desorption ionization time of flight (MALDI-TOF).

• BMB Reports
• /
• 제41권8호
• /
• pp.597-603
• /
• 2008

Atopic dermatitis (AD) is a chronic inflammatory skin disease that induces changes in various inflammatory skin cells. The prevalence of AD is as high as 18% in some regions of the world, and is steadily rising. However, the pathophysiology of AD is poorly understood. To identify the proteins involved in AD pathogenesis, a comparative proteomic analysis of protein expression in peripheral blood mononuclear cells isolated from AD patients and healthy donors was conducted. Significant changes were observed in the expressions of fourteen proteins, including the vinculin, PITPNB, and Filamin A proteins. Among the proteins, $\alpha$-SNAP and FLNA decreased significantly, and PITPNB increased significantly in AD patients compared with control subjects; these findings were further confirmed by real-time PCR and Western blot analysis. The comparative proteome data may provide a valuable clue to further understand AD pathogenesis, and several differentially regulated proteins may be used as biomarkers for diagnosis and as target proteins for the development of novel drugs.

• KSBB Journal
• /
• 제22권6호
• /
• pp.393-400
• /
• 2007

현재 많은 대학과 기업에서 다양한 방법으로 상용화가 가능한 protein microarray의 개발을 위해 많은 연구를 집중하고 있다. Protein microarray의 제작 및 분석 조건을 최적화하기 위한 연구도 진행되고 있지만 protein microarray로 부터 얻은 분석 결과를 모든 연구자들이 공유하고 통합하기 위한 노력이 절실한 실정이다. 뿐만 아니라, PCR 같은 무한 확장 방법이 존재하지 않는 단백질의 특성을 고려할 때, 좀 더 실용적인 protein microarray를 많이 만들기 위해서는 수많은 단백질들과 결합할 수 있는 특이성이 높고 결합력이 강한 capture molecule들을 개발하는 것이 필수이다. 그러나 이러한 장애에도 불구하고 protein microarray는 아주 적은 시료량으로 high-throughput assay가 가능하다는 장점 때문에 현재의 생명과학의 발전 추세로 볼 때 앞으로 protein microarray가 조만간 실용화될 것이며 이의 시장성은 매우 클 것으로 기대된다. 보다 빠른 실용화를 위해서는 protein microarray의 개발에 필요한 기반 기술의 개발과 동시에 이를 활용하기 위한 contents의 개발도 절실히 요구된다.

• 한국작물학회지
• /
• 제51권1호
• /
• pp.66-72
• /
• 2006

Spikelet proteins expressed at the young microspore stage in rice were separated and analysed by two-dimensional polyacrylamide gel electrophoresis (2DE). The separated proteins were electro blotted onto a polyvinylidene difluoride (PVDF) membrane, and 50 proteins were analyzed by a gas-phase protein sequencer. The N-terminal amino acid sequences of 20 out of 50 proteins were determined. N-terminal regions of the remaining proteins could not be sequenced because of blocking. The internal amino acid sequences of proteins were determined by sequence analysis of peptides obtained by the Cleveland peptide mapping method. Results revealed the presence of the photosynthetic apparatus at rice young microspore stage. Major proteins identified in this study could be used as a marker for various studies on physiological stresses.

• 한국환경농학회지
• /
• 제30권4호
• /
• pp.395-401
• /
• 2011

BACKGROUND: Potassium (K) is one of the macronutrients which are essential for plant growth and development. Its deficiency in paddy soils is becoming one of the limiting factors for increasing rice yield in Asia. METHODS AND RESULTS: To investigate physiological symptoms under K-starvation (NP) compared with complete media (NPK) condition, we measured shoot/root length, weight, nutrients, and patterns of protein expression. The shoot growth was significantly reduced, but root growth was not affected by K-starvation. However, biomasses were decreased in both shoot and root. Uptake of K was reduced up to 85%, while total concentrations of P, Ca, Mg, Na were increased in root and shoot. To better understand the starved K mechanism of rice, comparative proteome analysis for proteins isolated from rice leaves was conducted using 2-DGE. Five spots of differentially expressed proteins were analyzed by MALDI-TOF MS. Analysis of these K-starvation response proteins suggested that they were involved in metabolism and defense. CONCLUSION(s): Physiological and 2-DGE based proteomics approach used in our study results in observation of morphology or nutrients change and identification of K-starvation responsive proteins in rice root. These results have important roles in maintaining nutrient homeostasis and would also be useful for further characterization of protein function in plant K nutrition.

• 한국산학기술학회논문지
• /
• 제9권3호
• /
• pp.800-806
• /
• 2008

자연발효된 청국장으로부터 혈전용해활성을 가지는 세균 Bacillus licheniformis HK-12를 농화분리하여, 배양기간 동안 생산된 혈전용해활성을 가지는 단백질에 대해 프로테옴 분석을 실시하였다. B. licheniformis HK-12를 액체 영양배지에 접종하여 얻어진 배양상등액을 피브린 평판법(fibrin plate method)을 사용하여 효소활성을 측정하였다. 그 결과, HK-12의 혈전용해 활성은 대조구인 plasmin보다 약 2.3배정도 높은 활성을 나타내었다. 효소는 배양상등액을 ammonium sulfate 침전, DEAE-cellulose chromatography, Sephadex chromatography 등을 수행하여 분리정제하였으며, 정제된 혈전용해효소의 분자량은 SDS-PAGE를 통해 약 23 kDa로 측정되었다. 배양시간에 따른 HK-6의 세포외 단백질의 변화를 2-D PAGE 분석을 통하여 분석하였다. 그 결과 36시간배양 후에 가장 현저하게 유도된 spot #1을 분리하였으며, MALDI-TOF MS를 이용하여 단백질 동정을 실시한 결과, 유도된 단백질의 아미노산 서열은 $^1EKKIEKYREEEORLK^{15}$으로서, serine protein kinase (PrkA) (AAU22526)로 확인되었다.

• Journal of Ginseng Research
• /
• 제43권1호
• /
• pp.143-153
• /
• 2019

Background: Ginseng is one of the well-known medicinal plants, exhibiting diverse medicinal effects. Its roots possess anticancer and antiaging properties and are being used in the medical systems of East Asian countries. It is grown in low-light and low-temperature conditions, and its growth is strongly inhibited at temperatures above $25^{\circ}C$. However, the molecular responses of ginseng to heat stress are currently poorly understood, especially at the protein level. Methods: We used a shotgun proteomics approach to investigate the effect of heat stress on ginseng leaves. We monitored their photosynthetic efficiency to confirm physiological responses to a high-temperature stress. Results: The results showed a reduction in photosynthetic efficiency on heat treatment ($35^{\circ}C$) starting at 48 h. Label-free quantitative proteome analysis led to the identification of 3,332 proteins, of which 847 were differentially modulated in response to heat stress. The MapMan analysis showed that the proteins with increased abundance were mainly associated with antioxidant and translation-regulating activities, whereas the proteins related to the receptor and structural-binding activities exhibited decreased abundance. Several other proteins including chaperones, G-proteins, calcium-signaling proteins, transcription factors, and transfer/carrier proteins were specifically downregulated. Conclusion: These results increase our understanding of heat stress responses in the leaves of ginseng at the protein level, for the first time providing a resource for the scientific community.