The Biochemistry and Structural Biology Lab.

 

               

樓國隆 博士

Kuo-Long Lou, Ph.D

 

 NTU Distinguished Research Chair Professor in Membrane Protein Structure and Function

台大膜蛋白結構與功能特聘研究講座

 

NTU Membrane Protein Structure and Function Core Laboratory and Research Groups

台大膜蛋白結構與功能核心實驗室及研究群

 

 

 1st. Formosan Symposium on Structural Biology of Membrane Protein and Biomembrane

第一屆台灣膜蛋白暨生物膜結構生物學研討會

 

Prof. Mirek Cygler 訪台演講公告

  

台大膜蛋白特聘研究講座2012年諾貝爾獎得主駐校講學研究活動

 

PPDock (Portal Patch Dock) web server

 

 

Joint Appointments: Institute of Biochemistry and Molecular Biology, Medical College / Institute of Biotechnology, Bio-Resources and Agriculture College / Center for Biotechnology

合聘單位:

醫學院生化分生所 / 生農學院生技所 / 生物技術研究中心

 

 

Personal Profile

個人資料

 

 

Current Position:

Associate Professor of Biochemistry and Structural Biology

Graduate Institute of Oral Biology, School of Dentistry, College of Medicine / Institute of Biochemistry and Molecular Biology, College of Medicine / Institute of Biotechnology, College of Bio-Resources and Agriculture / Center for biotechnology, National Taiwan University / R.O.C.

 

Office Tel: +886-2-23123456 ext. 66616

Office Fax: +886-2-23820685

E-Mail: kllou@ntu.edu.tw

 

Degrees:

1987            Department of Environmental Engineering National Cheng-Kung University/R.O.C. (B.S.)

1989                    Institute of Biological Chemistry National Cheng-Kung University/R.O.C. (M.S.)

1993                    Department of Structural Biology, Biozentrum University of Basel/ Switzerland (M.A.)

1996            Department of Structural Biology, Biozentrum University of Basel/S witzerland (Ph.D.)

 

Professional Experiences:

1991-1992       Institute of Biology III University of Freiburg/ Germany (T.A.)

1992-1996       Department of Structural Biology, Biozentrum University of Basel/ Switzerland (T.A.)

1996                Department of Structural Biology, Biozentrum University of Basel/ Switzerland (Postdoc)

1997-1999       Department of Medical Physiology and Biophysics/ Neurosciences Research Groups, Faculty of Medicine, University of Calgary/ Canada (Research Associate)

1999-2003       Graduate Institute of Oral Biology/ Medical College / National Taiwan University (Assistant Professor)

2002-                     Institute of Biochemistry and Molecular Biology / Medical College/ National Taiwan University (Joint Appointment)

2008-                             Institute of Biotechnology/ Bio-Resources and Agriculture College/ National Taiwan University (Joint Appointment)

2009-              Center for Biotechnology/ National Taiwan University (Joint Appointment)

 

The research of Dr. Kuo-Long Lou has been recently focused on the studies of structure and functional regulations of potassium channels. These studies are mainly based upon the structural analysis in combination with biochemical and electrophysiological methods applied to develop the gating mechanisms of potassium channels in nanoscales or in atomic levels. Meanwhile, through the approach via toxin-channel interactions, Dr. Lou has proposed several novel theoretical hypotheses and therefore expanded the collaborations with other institutions both inside and outside the university.

       Huang, P. T., Shiau Y. S. & Lou K. L.* (2007). The Interaction of Spider Gating Modifier Peptides with Voltage-Gated Potassium Channels. Toxicon. 49, 285-292.

 

The structure-functional study of potassium channel regulations. We combine biochemistry, structural biology, bioinformatics, and cellular electrophysiology to approach and to investigate the molecular mechanism of Kir channel regulations (ROMK1 and KATP) and Kv channel gating. (in collaboration with pharmacology department).

(A) Toxin-channel interactions. The hanatoxins and other toxins, are investigated with facilities of synchrotron radiations. (international collaborations with Academia Sinica, NSRRC, pharmacology, Bragg Institutes and Sidney Univ.) In addition, regarding the molecular mechanism of binding-induced inhibition of gating in voltage-dependent K+-channels by hanatoxin, molecular docking simulations revealed significant conformational change of S3C segment in the structure of S3C-hanatoxin complex, which has been comprehended as a reasonable factor resulting in the inhibition of channel activation not through blocking directly on the pore. Further experiments with biochemical and kinetic methods, e.g., stopped-flow technique and fluorescence titration, were used to verify such hypothesis.

Reference:

Lou et al. (2002) J. Mol. Recognit. 15, 175-179

Huang et al. (2002) Recept. Channels 8, 79-85

Shiau et al. (2002) J. Mol. Model. 8, 243-247

Lou et al. (2003) J. Mol. Recognit. 16, 392-395

Shiau et al. (2003) Chem. Res. Toxicol. 16, 1217-1225

Huang et al. (2007). Toxicon 49, 285-292.

 

 

 

 

 

(B) Expression, purification and crystallization of Kir6.2 and the intracellular domains of Kir1.1 channels.

Reference:

Lou et al. (2001) J. Mol. Model. 7, 20-25

Kung et al. (2008) J. Formos. Med. Assoc. 107, 600-608

Lee et al. (2008) J. Mol. Graph. Model. 27, 332-341

 

 

(C) Structural analysis of the unique insecticidal activity of VrD1 (VrCRP) targeting on potassium channels and the homoplasy evolutionary links between plant defensins and arthropod neurotoxins. Methodology used includes mutational/electrophysiological analyses, proteomics, SAXS, and SANS (for membrane interaction detection, see also below).The toxin-membrane interactions. A novel plant defensin VrD1 is applied in the search of membrane target(s), whereas the interactions between biomembranes and VrD1, as well as hanatoxins and other toxins, are investigated with facilities of synchrotron radiations. (international collaborations with Academia Sinica, NSRRC, pharmacology, Bragg Institutes and Sidney Univ.)

Reference:

Shiau et al. (2006) J. Mol. Recognit. 19, 441-450

 

 

(D) The functional roles of potassium channels on masticatory muscle fatigue.

Reference:

Shiau et al. (2003) J. Oral Rehabil. 30, 978-984.

 

 (E) Structural study of caries proteins. (in collaboration with microbiology and dentistry departments) This includes the 3-D structural analysis of S. mutans glucosyltransferases. Update results of putative but constructive structural evidence have demonstrated and implied the possible mechanism of the reduced GTF enzyme activities due to monoclonal antibody recognition.

Reference:

Tsai et al. (2000) FEMS Microbiol. Lett. 188, 75-79

Chia et al. (2003) J. Mol. Model. 9, 153-158

 

 

(F) Development of Software for virtual drug screening and structural studies. Practical software will be shortly free and online available for academic purposes.

 

(G) Crystal structural analysis for the interactions of (membrane) proteins involved in carcinogenesis mechanism induced by Helicobacter pylori (including SAXS). (in collaboration with biochemistry department)

 

(H) Structural and functional characterizations of haemostatic/thrombolytic proteins (in angiogenesis). This study includes the Ca2+-induced conformational change and the dimerization of human thrombomodulin domains and the formation of ternary complex of Streptokinase-plasmin-plasminogen. (international collaborations with NCKU, biochemistry department, medical technology dept., NSRRC, and Sidney Univ.) Expression, purification and crystallization of various TM domains are also underway.

 

 

(I) Development of crystallization methods. (in collaboration with physics department)

 

 

現職:

生化暨結構生物學副教授

醫學院牙醫專業學院口腔生科所 / 醫學院生化分生所 / 生農學院生技所 / 生物技術研究中心

國立台灣大學 / 中華民國 台灣

 

Office Tel: +886-2-23123456 ext. 66616

Office Fax: +886-2-23820685

E-Mail: kllou@ntu.edu.tw

 

學歷:

1987            中華民國 台灣 國立成功大學環境工程學系 學士

1990                    中華民國 台灣 國立成功大學生物化所 碩士

1992                德國 福萊堡大學生物物理所 碩士

1994                    瑞士 巴塞爾大學結構生物學 / 生物中心 碩士

1996            瑞士 巴塞爾大學結構生物學 / 生物中心 博士士

 

經歷:

1991-1992       德國 弗萊堡大學 生物第三所生物物理講座 助教

1992-1996       瑞士 巴塞爾大學 結構生物學系/生物中心 助教

1996                瑞士 巴塞爾大學 結構生物學系/生物中心 博士後研究員

1997-1999       加拿大 卡加利大學 醫學生理及生物物理所 助研究員

1999-2003       國立台灣大學醫學院 口腔生科所 助理教授

2002-               國立台灣大學醫學院 生化分生所      合聘助理教授

2008-               國立台灣大學生農學院 生技所 合聘副教授

2009-       國立台灣大學生物技術研究中心 合聘副教授研究員

 

本人近五年來研究主要著重在人類齲齒蛋白結構及鉀離子通道之結構-功能調控。簡單的說,利用結構生物學之分析,配合生化及電生理的測試,在奈米或更微細的原子層面,深入了解分子反應之作用機轉。同時更透過toxin-channel interactions之研究,提出新的理論假說,也拓展了院內外的良好合作關係。

Huang, P. T., Shiau Y. S. & Lou K. L.* (2007). The Interaction of Spider Gating Modifier Peptides with Voltage-Gated Potassium Channels. Toxicon 49, 285-292.

 

研究題目則更擴及H. pylori GroES與人類胃壁上皮細胞TLR作用機制、血栓蛋白結構分析與篩藥軟體之開發等。

Shimamura et al. (2004) Structure 12, 1471-1480

Lin et al. (2007) Mol. and Cell. Proteomics 6, 1018-1026

Kim et al. (2007) Cell  130, 906-917

 

 

具體研究項目如下 (1)蜘蛛毒蛋白造成鉀離子通道抑制之分子機制。我們的研究已發現toxin 結合時透過pore以外的區域造成可能之形變,因而抑制通道開啟;也利用了生化合成及螢光標定,配合stopped-flow,以動力學分析來對上述假說進行實驗證明。此部份計發表了6SCI papers,詳見著作目錄。

參考文獻:

Lou et al. (2002) J. Mol. Recognit. 15, 175-179

Huang et al. (2002) Recept. Channels 8, 79-85

Shiau et al. (2002) J. Mol. Model. 8, 243-247

Lou et al. (2003) J. Mol. Recognit. 16, 392-395

Shiau et al. (2003) Chem. Res. Toxicol. 16, 1217-1225

Huang et al. (2007) Toxicon 49, 285-292.

 

 

 

(2)Kir1.1Kir6.2通道膜內區域之表現、純化及結晶。此部分與藥理科及藥學所合作。目的在透過晶體結構提供直接證據,來解釋無法用電生理清楚說明之gating機制。此部份已完成三篇SCI papers,正積極投稿中;其中一篇並已被接受發表在績優期刊上。

參考文獻:

Lou et al. (2001) J. Mol. Model. 7, 20-25

Kung et al. (2008) J. Formos. Med. Assoc. 107, 600-608

Lee et al. (2008) J. Mol. Graph. Model. 27, 332-341

 

 

(3) VrD1造成綠豆豆象幼蟲死亡之機制探討,由此發展至與鉀離子通道作用之電生理分析、蛋白質體學膜上標的物尋找、X-ray小角度散射結構分析(SAXS)、以及利用小角度中子散射結構分析(SANS)探討toxinsplasma membrane之反應細節。此部份計發表了1SCI paper (初步結構分析),第二篇(細胞電生理)亦接近完成。SAXSSANS的部份則獲得核准使用澳洲ANSTOBeamtimes

參考文獻:

Shiau et al. (2006) J. Mol. Recognit. 19, 441-450

 

 

(4)咀嚼肌疲乏與K+-channels關聯之研究。此部份計發表了1SCI paper

參考文獻:

Shiau et al. (2003) J. Oral Rehabil. 30, 978-984.

 

(5)齲齒蛋白GTFsstructure-function 之研究,由微生物所賈景山醫師initiated。目前已對單株抗體造成酵素活性改變之可能機制提出建設性的結構證據。此部份計發表了2SCI papers

參考文獻:

Tsai et al. (2000) FEMS Microbiol. Lett. 188, 75-79

Chia et al. (2003) J. Mol. Model. 9, 153-158

 

 

 

(6)篩藥結構軟體之開發。此部份計已完成了2SCI papers,計畫投往Nucleic Acids Research

 

 

(7)胃幽門螺旋桿菌(H. pylori)致癌機轉中膜蛋白作用之結構分析。此部份共同發表了1SCI paper,目前亦有突破性之 SAXS data

 

 

(8)血液調節素thrombomodulin在血管新生過程中之分子結構變化。為本計畫所提出之內容,有相當豐富之preliminary dataSAXSSANS的部份亦獲得核准使用澳洲ANSTOBeamtimes,因此為國合計劃。

 

 

(9)利用原子力顯微鏡開發理性結晶新方法,尚處於創始階段。