2011年6月15日 星期三

李展平老師實驗室介紹

專長:生物化學、分子生物學、微生物分子診斷學、奈米生物醫學

研究主題: 微生物分子診斷、奈米生物技術

說明:

1. Structure-Function of Nucleic Acids and Interactions between Nucleic Acid and Protein

One of our interests is to explore a very promising field of analyzing structural features of nucleic acids and nucleic acids-proteins interactions by molecular/biochemical analyses and direct image analyses using Scanning Probe Microscopy (SPM). We are in collaboration with Dr CC Wang’s group; they are specialized in the SPM technologies, at NCKU. Our molecular/ biochemical efforts are to clone the protein genes including RecA, SSB, T7 endonuclease I, pyrophosphatase, lambda integrase, P1 cre recombinase, Q Beta replicase, and many others, to express them in E. coli cells, to purify them with high purity and activity, and to analyze their functions in vitro and in vivo. DNA or RNA with specialized structural elements such as cruciform, H-DNA, attB/P site, pseudoknot will also be cloned or constructed. We expect these results (molecular/chemical and SPM) will provide us deeper insights into structural features of nucleic acids and even the dynamically structural changes under different conditions and also after interactions with proteins.

2. Molecular diagnosis of diseases caused by pathogens

For the past years, we have been focusing on the development of methods for the molecular diagnosis of diseases cause by pathogens, such as MTB, four blood-born viruses (HBV, HCV, HIV, and HTLV, assay performance see Figure 2) and many others. At the same period, we have also developed the method of using nano si-coated magnetic beads for the purification of viral DNA/RNA from clinical samples with low virus titer; cloned, expressed and purified PCR enzymes and related proteins to establish many PCR and RT-PCR assays. In addition, related technologies such as the use of nanogold for the detection of mutations and a new technology platform that would allow us to carry out high throughput screening of mutations and SNP, and also for genotyping analyses.

江信仲老師實驗室介紹

專長:細胞生物學、血管生物學、分子生物學

研究主題:1. CD93 對內皮細胞發炎的影響
2. 研究microRNA做為治療模式鼠視網膜新生血管病變潛在目標的
應用,以及對組織內部分子表達及新生血管形成的影響

說明:
血管內皮細胞(vascular endothelial cells)是位於血管最內層、與血液直接接觸的地方,具有調節血管緊張度、血管通透性及促進血管發生的作用,對於調控血管功能扮演重要角色,在調節血液流變性、血球細胞對血管壁粘附等方面也發揮關鍵作用。因此,內皮細胞功能障礙被認為是慢性血管併發症諸如糖尿病及心血管動脈硬化的關鍵因素。在正常情況下,白血球在血管中流動,並不會與血管內皮細胞黏著。但當體內有外來物入侵或發炎反應產生時,因組織受損,導致血管內皮細胞受到細胞刺激 (如TNF-α、LPS、IL-1、IL-6等)作用之下,影響血管內微環境的發炎反應。而發炎反應的發生乃藉由血管內皮細胞表現黏著分子(如ICAM-1及VCAM-1),促使循環白血球黏著到血管壁上,進一步移行到發炎組織部位,執行免疫反應。若血管內皮的屏障和通透性改變,使氧化低密度膽固醇增加,此變性的氧化低密度膽固醇,會刺激產生化學趨性物質,吸引血液中的單核球進入內皮層的內皮細胞下空隙,並轉變成巨噬細胞,將氧化低密度膽固醇吞噬,而反覆吞噬氧化低密度膽固醇的巨噬細胞,則終至變成泡沬細胞沈積在內皮層內。若此情形繼續進行,則一而再,再而三的巨噬細胞形成的泡沬細胞死亡而疊積,再加上結締組織增生與修補,則將造成早期的動脈硬化斑(fatty streak),若動脈硬化繼續進行,則形成動脈粥狀硬化塊(atherosclerotic plaque)。漸漸進展增大的動脈硬化塊,則會造成阻塞及狹窄之症狀,最終形成動脈硬化。CD93是一種表現在內皮細胞與單核球的C-type lectin 穿膜蛋白,之前我們的研究結果發現CD93會干擾血管內膽固醇的表現,在動物實驗中會促進血管不正常栓塞,因此研究CD93對刺激膽固醇表現的機制是本實驗室的主題之一。

早產兒視網膜病變(Retinopathy of Prematurity)是一種發生於早產兒或低體重新生兒的視網膜病變。主要是因為早產兒出生時視網膜尚未發育完全, 加上出生後在保溫箱內受到高濃度氧氣的刺激,先天不良與後天失調,造成視網膜血液供應系統失衡。輕者局部視網膜缺氧,重者大量不正常血管滋生,造成出血,甚至視網膜剝離,導至失明。血管新生(angiogenesis)是正常生理變化中(如生長、傷口癒合)所必需有的過程,近年來科學家們也發現它和癌症的發展有密切的關係。當周圍的結締組織刺激後會分泌許多促使血管新生的物質(angiogenic substances)激活血管內皮細胞,而發生下列變化:1) 腫瘤周圍結締組織的分解破壞; 2) 內皮細胞增生; 3) 內皮細胞向分泌促使血管新生物質的地方移動; 4) 內皮細胞重新組合成新生血管。本實驗室另一方向在研究高濃度氧氣環境下,內皮細胞被誘導產生血管新生,用來做為病變致病機轉的研究。

最近發表之論文:
1. Liu SL, Li YH, Shi GY, Tang SH, Jiang SJ, Huang CW, Liu PY, Hong JS, Wu HL. 2009 Apr. Dextromethorphan reduces oxidative stress and inhibits atherosclerosis and neointima formation in mice. Cardiovasc Res. 82(1):161-169. (SCI)
2. Jiang SJ, Campbell LA, Berry MW, Rosenfeld ME, Kuo CC. 2008 Jul. Retinoic acid prevents Chlamydia pneumoniae induced foam cell development in a mouse model of atherosclerosis. Microbes Infect. 10(12-13):1393-1397. (SCI)
3. Jiang SJ, Kuo CC, Berry MW, Lee AW, Campbell LA. 2008 Apr. Identification and characterization of Chlamydia pneumoniae specific antigens which activateF production in RAW 264.7 murine macrophages. Infect. Immun. 76(4): 1558-1564. (SCI)
4. Kuo CC, Lee A, Jiang SJ, Yaraei K, Campbell LA. 2007 Jul. Inoculation of Chlamydia pneumoniae or Chlamydia trachomatis with ligand that inhibit attachment to host cells reduces infectivity in the mouse model of lung infection: implication for anti-adhesive therapy. Microbes Infect. 9(9):1139-1141. (SCI)
5. Jiang SJ, Lin TM, Shi GY, Eng HL, Chen HY, Wu HL. 2004 Sep. Inhibition of bovine herpesvirus-4 replication in endothelial cells by arsenite. Antiviral Res. 63(3):167-175. (SCI)
6. Jiang SJ, Lin TM, Shi GY, Eng HL, Chen HY, Wu HL. 2004 Jul-Aug. Inhibition of bovine herpesvirus-4 replication by arsenite through downregulation of the extracellular signal-regulated kinase signaling pathway. J Biomed Sci. 11(4):500-510. (SCI)

劉哲文老師實驗室介紹

專長:生物化學、結構生物學、奈米生物醫學、蛋白質化學

研究主題及說明:

1. C型肝炎病毒(hepatitis C virus, HCV)核心蛋白之組裝及其與外套膜、核酸之交互作用. C型肝炎為目前造成慢性肝炎、肝硬化、甚至肝癌之主要原因之ㄧ。然而不同於B型肝炎,目前並沒有有效之疫苗,以及專一性之C型肝炎治療方法。主要的原因,即為C型肝炎病毒之培養非常困難,以致於對其之研究不易而了解不足。本實驗室針對組裝病毒核心顆粒之核心蛋白進行發酵槽之大量表達,研究其核心組裝,以期找出影響病毒核心組裝之關鍵,而對未來C型肝炎之治療做出貢獻。本實驗室並研究脂質以及RNA對於HCV核心蛋白結構之影響,以了解核心內之物質與核心外物質在核心組裝扮演之角色。

2. 原子力顯微鏡(atomic force microscopy)生物奈米影像分析及蛋白質結構分析技術. 原子力顯微鏡為現今奈米科技最重要之工具之ㄧ,本實驗室使用目前台灣最先進之生物用原子力顯微鏡,並配合電子顯微鏡及共軛焦顯微鏡等影像技術來對生物樣本進行奈米等及解析度之分析。本實驗室使用生物資訊結構分析法傅立葉轉換紅外光光譜儀(FTIR)並定期至新竹國家同步輻射研究中心(NSRRC)使用同步輻射圓二色光譜儀(SRCD)、小角度X光繞射儀(small angle X ray)等國家級貴重儀器對蛋白質結構進行分析。

3. 抗菌胜肽(antibacterial peptide)之生物資訊學設計、合成與殺菌作用專一性之研究. 由於近年來抗生素之濫用,使得多重抗藥性菌株不斷之產生,造成醫療上非常嚴重的問題,尋找新的抗生素便成為目前之重要課題之ㄧ。抗菌胜肽為目前產業界非常熱門之領域。本實驗室使用系統性、高效率之生物資訊學對抗菌胜肽進行設計、突變、及篩選,再利用目前產業界所使用之化學合成法對所選出之抗菌胜肽進行合成,並探討胜肽結構對細菌之殺菌效果,及哺乳動物細胞之選擇性關係,將會是產業界非常受歡迎之技術。

最近發表之論文:
1. Liou JW, Gu MH, Chen YK, Chen WY, Chen YC, Tseng YH, Hung YJ, Chang HH. Visible Light Responsive Photocatalyst Induces Progressive and Apical-Terminus Preferential Damages on Escherichia coli Surfaces. PLoS One. 2011. 6(5):e19982.
2. Kumar VB, Yuan TC, Liou JW, Yang CJ, Sung PJ, Weng CF. Antroquinonol inhibits NSCLC proliferation by altering PI3K/mTOR proteins and miRNA expression profiles. Mutat Res. 2011. 707(1-2):42-52.
3. Chan SC, Lo SY, Liou JW, Lin MC, Syu CL, Lai MJ, Chen YC, Li HC. Visualization of the structures of the hepatitis C virus replication complex. Biochem Biophys Res Commun. 2011. 404(1):574-8.
4. Liou JW, Mulet X, Klug DR. Absolute measurement of phosphorylation levels in a biological membrane using atomic force microscopy: the creation of phosphorylation maps. Biochemistry. 2002. 41(27):8535-9.

李惠春老師實驗室介紹

專長:蛋白質化學、蛋白質結構與功能

研究主題: 探討C型肝炎病毒套膜蛋白(E1、E2)結構與功能之關係

說明:
許多動物病毒外都包覆了一層由宿主細胞膜所衍生的膜,膜上分佈有病毒的醣蛋白,有些外套膜上的醣蛋白協助辨識宿主細胞。當病毒被宿主細胞吞噬初期會形成vesicle,病毒的外套膜便會與vesicle的膜融合,而將病毒的遺傳物質釋放至宿主細胞的細胞質中。此膜融合的過程便是由病毒套膜上的醣蛋白所負責的,C肝病毒的E1及E2蛋白疑似具有這樣的功能。目前,C肝病毒的E1及E2蛋白被歸類為第二型膜融合蛋白,即先以dimmers型式存在,受酸鹼值改變而形成trimer,再藉由構型改變使兩層膜融合為一。只是在C肝病毒中,此過程的分子機制及蛋白質構形變化尚不明瞭。

我們利用E. coli 大量表達並純化C肝病毒的E1及E2蛋白,配合生化及生物物理的方法來探討它們在膜融合過程中的角色,尤其是參與膜融合的序列、在膜融合前與融合後的蛋白構形、及因酸鹼值改變所引起的構形改變。

最近發表之論文:
1. Bertrand Vileno, Jean Chamouna, Hua Liang, Paul Brewerc, Brian D. Haldemanc, Kevin C. Facemyer, Bridget Salzameda, Likai Song, Hui-Chun Li, Christine R. Cremo, and Piotr G. Fajer. 2011. Broad disorder and the allosteric mechanism of myosin II regulation by phosphorylation. Proc Natl Acad Sci USA. 108:8218-23.

2. Shih-Ching Chan, Shih-Yen Lo, Je-Wen Liou, Min-Ching Lin, Ciao-Ling Syu, Meng-Jiun Lai , Yi- Cheng Chen, Hui-chun Li. 2011. Visualization of the structures of the hepatitis C virus replication complex. Biochem. Bioph. Res. Co. 404:574-578.

3. Chee-Hing Yang, Hui-Chun Li, Jeng-Geng Jiang, Che-Fang Hsu, Yi-Jen Wang, Meng-Jiun Lai, Yue-Li Juang and Shih-Yen Lo. 2010. Enterovirus type 71 2A protease functions as a transcriptional activator in yeast. J. Biomed Sci. 17:65.

4. Chia-Wei Chang, Hui-Chun Li, Che-Fang Hsu, Chiao-Yen Chang and Shih-Yen Lo. 2009. Increased ATP generation in the host cell is required for efficient vaccinia virus production. J. Biomed Sci. 16:80-89.

5. Shih-Chi Chen, Shih-Yen, Lo, Hsin-Chieh, Ma, Hui-Chun Li. 2009. Expression and membrane integration of SARS-CoV E protein and its interaction with M protein. Virus Genes 38:365-371.

6. Yi-Ching Hsieh, Hui-Chun Li, Shih-Chi Chen and Shih-Yen Lo. 2008. Interactions between M protein and other structural proteins of severe, acute respiratory syndrome-associated coronavirus. J. Biomed Sci. 15:707-717.

7. Hsin-Chieh Ma, Ta-Wei Lin, Hui-Chun Li, Sanae M.M. Iguchi-Ariga, Hiroyoshi Ariga, Yu-Li Chuang, Jing-Hsiung Ou, Shih-Yen. 2008. Hepatitis C virus ARFP/F protein interacts with cellular MM-1 protein and enhances the gene trans-activation activity of c-Myc. J. Biomed Sci. 15:417-425.

8. Hsin-Chieh Ma, Chiu-Ping Fang, Yi-Ching Hsieh, Shih-Chi Chen, Hui-Chun Li, Shih-Yen Lo. 2008. Expression and membrane integration of SARS-CoV M protein. J. Biomed Sci. 15:301-310.

9. Li, H.-C., Song, L., Salzameda, B., Cremo, C. R. Fajer, P. G. 2006. Rugulatory and Catalytic Domain Dynamics of Smooth Muscle Myosin Filaments. Biochemistry 45;6212-6221.

陳灝平老師實驗室介紹

專長:微生物基因體學、蛋白質化學、酵素與輔酶化學、化學生物學

研究方向:
 
1.基因體學與轉錄體學在臨床醫學的應用
2.B12代謝異常個案在分子醫學層次的探討
3.酵素活性分析
4.臨床檢驗方法的改進與開發
5.開發新型生物能源與光電材料


最近發表之論文:
1. HP Chen, M Chow, CC Liu, Al Lau, J Liu, and LD Eltis,* "Vanillin and vanillate catabolism in Rhodococcus jostil RHA1." (2011) (manuscript in preparation)

2. HP Chen, S Zhu, A Lau, AE Davis, SWM Eng, CG Howes, FH Crocker, KJ Indest, SJ Hallam, and LD Eltis,*"Complete genome sequence of Gordonia sp. KTR9." (2011) (manuscript in preparation)

3. KJ Indest,* CM Jung, HP Chen, D Hancock , C Florizone, LD Eltis,* FH Crocker “Functional Characterization of pGKT2, a 182-Kilobase Plasmid Containing the xplAB Genes, Which Are Involved in the Degradation of Hexahydro-1,3,5-Trinitro-1,3,5-Triazine by Gordonia sp. Strain KTR9.”(2010) Applied and Environmental Microbiology 76, 6329-6337.

4. CH Hsu,* and HP Chen* “Molecular modeling studies of the conserved B12-binding motif and its variants from Clostridium tetanomorphum glutamate mutase” (2010) Protein & Peptide Letters 17 759-764.

5. HP Chen,* HJ Hsu, FC Hsu, CC Lai, and CH Hsu, “The interactions between coenzyme B12 analogs and adenosylcobalamin-dependent glutamate mutase from Clostridium tetanomorphum.” (2008) FEBS Journal 275, 5960-5968.

6. CH Tseng, CH Yang, HJ Lin, C Wu, and HP Chen,* “The S subunit of D-ornithine aminomutase from Clostridium sticklandii is responsible for the allosteric regulation in D-alpha-lysine aminomutase.” (2007) FEMS Microbiology Letters 274,148-153.

7. YP Weng, WS Yang, FC Hsu, and HP Chen,* “Optimization of the overexpression of glutamate mutase S component under the control of T7 system by using lactose and IPTG as the inducers” (2006) Enzyme and Microbial Technology 38, 465-469.

8. FC Hsu, TJ Ho, CC Lai, CF Lin, and HP Chen,* “Cloning, sequencing, expression, and single-step purification of the adenosylcobinamide kinase/ adenosylcobinamide-phosphate guanylyltransferase (CobU) from Salmonella typhimurium ATCC 19585” (2005) Protein Expression and Purification, 42, 178-181.

9. HP Chen,* FC Hsui, LY Lin, CT Ren, and SH Wu, “Coexpression, purification, and characterization of the E and S subunits of coenzyme B12 and B6 dependent Clostridium sticklandii D-ornithine aminomutase in Escherichia coli” (2004) European Journal of Biochemistry, 271, 4293-4297.

10. HP Chen,* FD Lung, CC Yeh, HL Chen, and SH Wu, “The role of the conserved histidine-aspartate pair in the ‘base-off’ binding of cobalamins”. (2004) Bioorganic and Medicinal Chemistry, 12, 577-582.

11. HP Chen,* SH Wu, YL Lin, CM Chen and SS Tsay, “Cloning, sequencing, heterologous expression, purification and characterization of adenosylcobalamin-dependent D-ornithine aminomutase from Clostridium sticklandii”. (2001) Journal of Biological Chemistry, 276, 44744-44750.

12. HP Chen,* CF Lin, YJ Lee, SS Tsay and SH Wu, “Purification and properties of ornithine racemase from Clostridium sticklandii” (2000) Journal of Bacteriology, 182, 2052-2054.