Proteomic Analysis기법을 이용한 홍화자약침액(紅花子藥鍼液)이 간암세포주(肝巖細胞柱)의 유전자(遺傳子) 발현(發顯)에 미치는 영향(影響)

이경민,임성철,정태영,서정철,한상원,Lee, Kyung-Min,Lim, Seong-Chul,Jeong, Tae-Young,Seo, Jung-Chul,Han, Sang-Won 대한약침학회 2005 Journal of pharmacopuncture Vol.8 No.2

Objective : It has long been known about the osteogenic effect of CTF-HAS on bone tissues. However, it has not been determined the effect of CTF-HAS on cancer cells. The purpose of this study is to screen the CTF-HAS mediated differentially expressed genes in cancer cells such as HepG2 hepatoma cells lines. Methods : CTF-HAS was prepared by boiling and stored at $-70^{\circ}C$ until use. For proteomic analysis, total protein was analyzed by 2D gel electrophoresis and Q-TOF mass spectrometer. Results : In proteomic analysis, three spots were identified by 2D-gel electrophoresis and Q-TOF analysis. One down-regulated protein was heat shock 70kDa protein 5 and up-regulated proteins were chaperonin and 2-phospho -pyruvate-hydratase ${\alpha}-enolase$ by 1.5mg/ml of CTF-HAS. Discussion : Proteomic analysis approach were performed to screen the differential expression genes. The screened genes will be used for the better understanding in therapeutic effect of CTF-HAS on cancer field.

Salmonella Gallinarum 세포외막단백질의 프로테옴 분석 및 닭에서의 방어능 효과

선지선,조영재,장주현,강정무,한장혁,한태욱,Sun, Jisun,Cho, Youngjae,Jang, Joo-Hyun,Kang, Zheng-Wu,Han, Jang-Hyuk,Hahn, Tae-Wook 한국축산식품학회 2013 한국축산식품학회지 Vol.33 No.2

Salmonella Gallinarum (SG) is known as an important pathogen that causes fowl typhoid in chickens. To investigate SG outer-membrane proteins (OMPs) as a vaccine candidate, we used proteomic mapping and database analysis techniques with extracted OMPs. Also, extracted OMPs were evaluated in several aspects to their safety, immune response in their host and protective effects. Our research has established a proteomic map and database of immunogenic SG-OMPs used as inactive vaccine against salmonellosis in chickens. A total of 22 spots were detected by 2-dimensional gel electrophoresis and immunogenic protein analysis. Eight spots were identified by Matrix-Assisted Laser Desorption/Ionization-Time of Flight-Mass spectrometry (MALDI-TOF-MS) and peptide mass fingerprinting (PMF) and categorized into four different types of proteins. Among these proteins, OmpA is considered to be an immunogenic protein and involved in the hosts' immune system. To estimate the minimum safety dose in chickens, 35 brown layers were immunized with various concentrations of OMPs, respectively. Consequently, all chickens immunized with more than a $50{\mu}g$ dose were protected against challenges. Moreover, intramuscular administration of OMPs to chickens was more effective compared to subcutaneous administration. These results suggest that the adjuvanted SG-OMP vaccine not only induces both the humoral and cellular immune response in the host but also highly protects the hosts' exposed to virulent SG with $50{\mu}g$ OMPs extracted by our method.

Proteome analysis of sera of patients with active Behcet`s disease

( Zhenlong Zheng ),( Young In Lee ),( Jae Hee Cheon ),( Dongsik Bang ),( Do Young Kim ) 대한피부과학회 2015 대한피부과학회 학술발표대회집 Vol.67 No.2

Background: Behcet’s disease (BD) is a chronic, multisystemic vasculitis that theoretically affects all sizes and types of blood vessels. The validated biological marker to determine the disease activity of BD is still lacking. Therefore, our aim of this study was to search relevant markers for active BD using proteomic analysis. Objectives: The aim of this study was to search relevant markers for active BD using proteomic analysis. Methods: To find a novel serum marker in BD, we used pooled sera (equal volumes from 5 active BD patients and 5 healthy volunteer donors) and proteomics approach. For proteomics analysis, two-dimensional gel electrophoresis along with matrix-assisted laser desorption ionization time-of-flight mass spectrometry were performed. Results: There was significant differential expression of proteins in active BD and healthy control. We identified 13 maximally differentially expressed proteins after LC-MALDI-TOF/TOF analysis of 21 spots. Among 13 proteins, four (visual system homeobox 2, complement receptor Cr2, complement component C3c, truncated human apolipoprotein A) was downregulated and two proteins (Haptoglobin, FAM123A protein) were up-regulated in BD compared with healthy control. In addition seven proteins (myosin-15 precursor, vitronectin, heparin cofactor Ii, growth-inhibiting protein 25, bamacan homolog, amyloid related serum protein SAA, proapolipoprotein) were only detected in BD serum. Conclusion: Our study provides potential protein markers of active BD.

Pressure Cycling Technology-assisted Protein Digestion for Efficient Proteomic Analysis

Hyunsu Choi,Sang Kwang Lee,Kyung-Hoon Kwon,Jong Shin Yoo,김진영,Kelly Ji 대한화학회 2011 Bulletin of the Korean Chemical Society Vol.32 No.2

In typical proteomic analysis, trypsin digestion is one of the most time-consuming steps. Conventional proteomic sample preparation methods use an overnight trypsin digestion method. In this study, we compared high-pressure cycling technology (PCT) during enzyme digestion for proteome analysis to the conventional method. We examined the effect of PCT on enzyme activity at temperatures of 25, 37, and 50 ^oC. Although a fast digestion (1 h) was used for the standard protein mixture analysis, the PCT-assisted method with urea showed better results for protein sequence coverage and the number of peptides identified compared with the conventional method. There was no significant difference between temperatures for PCT-assisted digestion; however, we selected PCT-assisted digestion with urea at 25 ^oC as an optimized method for fast enzyme digestion, based on peptide carbamylation at these conditions. The optimized method was used for stem cell proteome analysis. We identified 233, 264 and 137 proteins using the conventional method with urea at 37 ^oC for 16h, the PCT-assisted digestion with urea at 25 ^oC for 1 h, and the non-PCT-assisted digestion with urea at 25 ^oC for 1 h, respectively. A comparison of these results suggests that PCT enhanced the enzyme digestion by permitting better access to cleavage sites on the proteins.

Pressure Cycling Technology-assisted Protein Digestion for Efficient Proteomic Analysis

Choi, Hyun-Su,Lee, Sang-Kwang,Kwon, Kyung-Hoon,Yoo, Jong-Shin,Ji, Kelly,Kim, Jin-Young Korean Chemical Society 2011 Bulletin of the Korean Chemical Society Vol.32 No.2

In typical proteomic analysis, trypsin digestion is one of the most time-consuming steps. Conventional proteomic sample preparation methods use an overnight trypsin digestion method. In this study, we compared high-pressure cycling technology (PCT) during enzyme digestion for proteome analysis to the conventional method. We examined the effect of PCT on enzyme activity at temperatures of 25, 37, and $50^{\circ}C$. Although a fast digestion (1 h) was used for the standard protein mixture analysis, the PCT-assisted method with urea showed better results for protein sequence coverage and the number of peptides identified compared with the conventional method. There was no significant difference between temperatures for PCT-assisted digestion; however, we selected PCT-assisted digestion with urea at $25^{\circ}C$ as an optimized method for fast enzyme digestion, based on peptide carbamylation at these conditions. The optimized method was used for stem cell proteome analysis. We identified 233, 264 and 137 proteins using the conventional method with urea at $37^{\circ}C$ for 16h, the PCT-assisted digestion with urea at $25^{\circ}C$ for 1 h, and the non-PCT-assisted digestion with urea at $25^{\circ}C$ for 1 h, respectively. A comparison of these results suggests that PCT enhanced the enzyme digestion by permitting better access to cleavage sites on the proteins.

프로테오믹 기법을 이용한 위암 바이오마커 연구

정상호(Sang-Ho Jeong),이영준(Young-Joon Lee),홍순찬(Soon-Chan Hong),하우송(Woo-Song Ha) 대한종양외과학회 2016 Korean Journal of Clinical Oncology Vol.12 No.1

Rapid Identification of ${\delta}-endotoxin$ from Bacillus thuringiensis subsp. kurstaki HD-1 with Proteomic Analysis

Lee, Dae-Weon,Je, Yeon-Ho,Koh, Young-Ho Korean Society of Applied Entomology 2006 Journal of Asia-Pacific Entomology Vol.9 No.4

This study was carried out to identify rapidly ${\delta}-endotoxin$ from Bacillus thuringiensis (Bt) subsp. kurstaki HD-1 with proteomic analysis. Protoxin was isolated from sporulated cells and purified by ultracentrifugation using 40-70% sucrose density gradient. Protoxin was treated with trypsin to analyze digested peptides by liquid chromatography-tandem mass spectrometry. The proteomic analysis for detected peptides revealed that this methodology is available for discriminating similar Bt strains by identifying Bt subsp. kurstaki HD-1-specific peptides, suggesting that proteomic analysis can be used for rapid identification of Bt strains.

Proteomic pattern-based analyses of light responses in Arabidopsis thaliana wild-type and photoreceptor mutants

Kim, Dong Su,Cho, Dae Shik,Park, Won-Man,Na, Hyung Jin,Nam, Hong Gil WILEY-VCH 2006 Proteomics Vol. No.

<P>Light critically affects the physiology of plants. Using two-dimensional gel electrophoresis, we used a proteomics approach to analyze the responses of Arabidopsis thaliana to red (660 nm), far-red (730 nm) and blue (450 nm) light, which are utilized by type II and type I phytochromes, and blue light receptors, respectively. Under specific light treatments, the proteomic profiles of 49 protein spots exhibited over 1.8-fold difference in protein abundance, significant at p <0.05. Most of these proteins were metabolic enzymes, indicating metabolic changes induced by light of specific wavelengths. The differentially-expressed proteins formed seven clusters, reflecting co-regulation. We used the 49 differentially-regulated proteins as molecular markers for plant responses to light, and by developing a procedure that calculates the Pearson correlation distance of cluster-to-cluster similarity in expression changes, we assessed the proteome-based relatedness of light responses for wild-type and phytochrome mutant plants. Overall, this assessment was consistent with the known physiological responses of plants to light. However, we also observed a number of novel responses at the proteomic level, which were not predicted from known physiological changes.</P>

Proteome Analysis on Differentially Expressed Proteins of the Fat Body of Two Silkworm Breeds, Bombyx mori, Exposed to Heat Shock Exposure

S. Hossein Hosseini Moghaddam,Xin Du,Jun Li,Jinru Cao,Boxiong Zhong,YuYin Chen 한국생물공학회 2008 Biotechnology and Bioprocess Engineering Vol.13 No.5

Proteomes of heat tolerant (multivoltine) and heat susceptible (bivoltine) silkworms (Bombyx mori) in response to heat shock were studied. Detected proteins from fat body were identified by using MALDI-TOF/TOF spectrometer, MS/MS, and MS analysis. Eight proteins, including small heat shock proteins (sHSPs) and HSP70, were expressed similarly in both breeds, while 4 protein spots were expressed specifically in the bivoltine breed and 12 protein spots were ex-pressed specifically in the multivoltine breed. In the present proteomics approach, 5 separate spots of sHSP proteins (HSP19.9, HSP20.1, HSP20.4, HSP20.8, and HSP21.4) were identified. Protein spot intensity of sHSPs was lower in the multivoltine breed than in the bivoltine breed after the 45C heat shock treatment, while the difference between two breeds was not significant after the 41C heat shock treatment. These results indicated that some other mechanisms might be engaged in thermal tolerance of multivotine breed except for the expression of sHSP and HSP70. There were visible differences in the intensity of heat shock protein expression between male and female, however, differences were not statistically significant.

Post-translational Modifications and Their Biological Functions : Proteomic Analysis and Systematic Approaches

Seo, Jawon,Lee, Kong-Joo 한국생화학분자생물학회 2004 BMB Reports Vol.37 No.1

Recently produced information on post-translational modifications makes it possible to interpret their biological regulation with new insights. Various protein modifications finely tune the cellular functions of each protein. Understanding the relationship between post-translational modifications and functional changes ("post-translatomics") is another enormous project, not unlike the human genome project. Proteomics, combined with separation technology and mass spectrometry, makes it possible to dissect and characterize the individual parts of post-translational modifications and provide a systemic analysis. Systemic analysis of post-translational modifications in various signaling pathways has been applied to illustrate the kinetics of modifications. Availability will advance new technologies that improve sensitivity and peptide coverage. The progress of "post-translatomics", novel analytical technologies that are rapidly emerging, offer a great potential for determining the details of the modification sites.