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Transcript
Chow, Lu-Ping 周 綠 蘋 蛋白 體暨蛋白 質功能核心 實驗 室 Proteomics and Protein Function Core Laboratory 所長 ,臺 大 醫學 院 生物化 學 暨分 子生 物學 研究 所 N T U R e s e a rc h Ce n te r Fo r M e d i c a l E xce l l e n ce 14 EDUCATION AND POSITIONS HELD: • • • • • • • • B.D. Department of Botany, National Chung Hsing University, Taiwan, 1980 Master D. Institute of Plant Biology, National Taiwan University, Taiwan, 1983 Ph.D. Department of Pharmaceutical Sciences, Tokyo University of Science, Tokyo, 1996 Instructor, Institute of Biochemistry & Molecular Biology College of Medicine, National Taiwan University, 1987- 1996 Associate Professor, Institute of Biochemistry & Molecular Biology, College of Medicine, National Taiwan University, 1996- 2001 Vice director, Department of Medical Genetics, National Taiwan University Hospital, 2001- 2007 Professor, Institute of Biochemistry & Molecular Biology, College of Medicine, National Taiwan University, 2001- 2008 Professor and chairman, Institute of Biochemistry & Molecular Biology, College of Medicine, National Taiwan University, 2008/12- Present HONORS: • • • • Biochemical J. reviewer Proteomics reviewer J. Proteome Research reviewer FEBS J. reviewer RESEARCH INTERESTS: 1. Bacterial virulence factors and cancer development 2. Host-pathogen interaction and cancer biomarkers discovery 3. Biomarkers of gastric diseases 4. Hepatocellular carcinoma (HCC)-related serum biomarkers SELECTED PUBLICATIONS: 1. Chan CH, Ko CC, Chang JG, Chen SF, Wu MS, Lin JT, and Chow LP*. Subcellular and Functional Proteomic Analysis of the Cellular Responses Induced by Helicobacter pylori. Molecular & Cellular Proteomics 2006, 5(4) :702–13. 2. Lin YF, Wu MS, Chang CC, Lin SW, Lin JT, Sun YJ, Chen DS, and Chow LP*. Comparative immunoproteomics of identification and characterization of virulence factors from Helicobacter pylori related to gastric cancer. Molecular & Cellular Proteomics 2006, 5(8) :1484-96. 3. Lin YF, Chen CY, Tsai MH, Wu MS, Wang YC, Chuang EY, Lin JT, Yang PC, and Chow LP*. Duodenal ulcer-related antigens from Helicobacter pylori: immunoproteome and protein microarray approaches. Molecular & Cellular Proteomic s 2007, 6(6) : 1018-26. 4. Chiu LL, Lee KL, Lin YF, Chu CY, Su SN, and Chow LP*. Secretome analysis of novel IgE-binding proteins from Penicillium citrinum. Proteomics-clinical applications 2008, 2(1): 33-45. 5. Wu MS, Chow LP*, Lin JT, and Chiou SH. Proteomic identification of biomarkers related to Helicobacter pylori-associated gastroduodenal disease: challenges and opportunities. J Gastroenterol Hepatol . 2008, 23(11) : 1657-61. 蛋白體暨蛋白質功能核心實驗室 Proteomics and Protein Function Core Laboratory Proteomics, a subset of functional genomics, is the study of the proteins expressed by an organism. These studies link genome sequence data and gene function. More than 200 proteomics facilities have been established worldwide. As a leading academic institution in Taiwan, National Taiwan University's core of services can provide fast, high-sensitivity identification of proteins by mass spectrometry performed with two instruments. MALDI-TOF (Matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry identifies proteins by peptide mass fingerprinting. Liquid chromatography-tandem mass spectrometry (using Q-STAR) enables researchers to analyze peptide sequence and post-translational modification, using monosaccharide analysis, oligosaccharide profiling, glycosylation sites and detection of phosphorylated amino acids. Furthermore, the facility offers consultation and support for protein separation techniques, proteome -related bioinformatics and data management. Our ongoing projects are designed to identify all the components of cancer and infectious disease proteomes, providing a basis for functional studies. We will also address identification of functional components of selected subcellular components. The overarching goal of achieving new understanding of diseases should yield important new targets for therapeutic interpretation and markers for early detection as well. The stated mission of the Proteome Core Facility is to provide comprehensive technological support for biomedical research, both basic and with potential clinical applications, on topics ranging from cancer and pathogenesis of infectious diseases to drug and vaccine design. Our services include: 1. Two-dimensional electrophoresis (2-DE) analysis (CBB, Silver Stain, Ruby, Cy3/Cy5 labling) 2. Protein digestion (In gel, In solution) 3. Identification of protein/peptide molecular weight by mass spectrometry (MALDI-TOF) 4. Identification of peptide sequence and posttranslational modification (LC-MS/MS) 5. Prediction of unknown protein by mass spectrometry database search with MASCOT software (MALDI-TOF, LC-MS/MS) │ Proteomics and Protein Function Core Laboratory The completion of the genomes for a large number of organisms has begun to revolutionize experimental approaches to a variety of problems in biology and medicine. The proteome is of intense interest to investigators because proteins are the major functional components of the cell. In the era of genomic research, characterization of a protein links it directly to its corresponding gene; understanding the function of a particular protein leads directly to gene function. Lu-Ping Chow, Ph.D 蛋白體暨蛋白質功能核心實驗室 教授 周綠蘋 15 -d im en sio n IE F1 N T U R e s e a rc h Ce n te r Fo r M e d i c a l E xce l l e n ce PA G E im en sio n Figure 1. Proteomics two-dimensional electrophoresis (2-DE) equipments (A) (B) (D) 16 2-d (C) (E) Figure 2. Hardware equipments and core services. (A) Fluorescence Microscopy; (B) Akta FPLC purification system; (C) Typhoon 9400 scanner; (D) MALDI-TOF mass spectrometry; (E) LC-MS/MS. 蛋白體暨蛋白質功能核心實驗室 Strategies for glycoprotein identification and characterization → → → → → → Figure 4. Strategies for glycoprotein identification and characterization. │ Proteomics and Protein Function Core Laboratory Figure 3. Strategy of protein profile alteration between normal and abnormal cells. 17 N T U R e s e a rc h Ce n te r Fo r M e d i c a l E xce l l e n ce 申請服務流程 Cells (state B) Cells (state A) Heavy AA Light AA SILAC labeling 選擇特定的服務細項<二維電泳分析、 蛋白質水解分析、微量蛋白質定序(LCMS/MS)、基質輔助雷射脫附游離/飛行 時間質譜(MALDI-TOF MS)>下載其申請表 格,來電預約後,將填妥之申請表與檢 體一併親自送至蛋白質暨蛋白質功能核 心實驗室。 Tryptic digestion of cellular proteins Phosphopeptide enrichment 初步核定分析樣品之品質與申請表格填 寫之完整度後,依收件順序進行測試。 LC-MS/MS MASCOT analysis Ingenuity Pathways Analysis Figure 5. Strategies for phosphoprotein identification and characterization. 18 從台大醫學院卓越中心的網路首頁中, 選擇「蛋白質暨功能」的分項子計畫進 入,再點選左側「申請服務」之選項。 實驗完成後,將以e-mail 寄發結果或通 知使用者領取分析結果,並依收費標準 收費<網頁有收費標準可供參考>。 Contact information 信箱 : [email protected] 聯絡人: 周綠蘋 聯絡分機 : 88682