請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61334
完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 楊啟伸 | |
dc.contributor.author | Ming-Jin Jheng | en |
dc.contributor.author | 鄭明晉 | zh_TW |
dc.date.accessioned | 2021-06-16T13:01:13Z | - |
dc.date.available | 2013-08-09 | |
dc.date.copyright | 2013-08-09 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-07 | |
dc.identifier.citation | 1 Rao, V. R. & Oprian, D. D. Activating mutations of rhodopsin and other G protein-coupled receptors. Annual review of biophysics and biomolecular structure 25, 287-314 (1996).
2 Spudich, J. L., Yang, C.-S., Jung, K.-H. & Spudich, E. N. Retinylidene proteins: structures and functions from archaea to humans. Annual review of cell and developmental biology 16, 365-392 (2000). 3 Briggs, W. R. & Spudich, J. L. Handbook of photosensory receptors. (Wiley-VCH, 2005). 4 Oesterhelt, D. & Stoeckenius, W. Functions of a new photoreceptor membrane. Proceedings of the National Academy of Sciences 70, 2853-2857 (1973). 5 Matsuno-Yagi, A. & Mukohata, Y. Two possible roles of bacteriorhodopsin; a comparative study of strains of Halobacterium halobium differing in pigmentation. Biochemical and biophysical research communications 78, 237-243 (1977). 6 Bogomolni, R. A. & Spudich, J. L. Identification of a third rhodopsin-like pigment in phototactic Halobacterium halobium. Proceedings of the National Academy of Sciences 79, 6250-6254 (1982). 7 Takahashi, T., Mochizuki, Y., Kamo, N. & Kobatake, Y. Evidence that the long-lifetime photointermediate of s-rhodopsin is a receptor for negative phototaxis in Halobacterium halobium. Biochemical and biophysical research communications 127, 99-105 (1985). 8 Hoff, W. D., Jung, K.-H. & Spudich, J. L. Molecular mechanism of photosignaling by archaeal sensory rhodopsins. Annual review of biophysics and biomolecular structure 26, 223-258 (1997). 9 Spudich, J. L. The multitalented microbial sensory rhodopsins. Trends in microbiology 14, 480-487 (2006). 10 林修平. 以嵌合蛋白質探討 HmBR I 第三胞內環之功能. 國立台灣大學碩士論文 (2010). 11 Oesterhelt, D. & Stoeckenius, W. Rhodopsin-like protein from the purple membrane of Halobacterium halobium. Nature 233, 149-152 (1971). 12 Oesterhelt, D. & Stoeckenius, W. Isolation of the cell membrane of Halobacterium halobium and its fractionation into red and purple membrane. Methods Enzymol. 31, 667-678 (1974). 13 Oesterhelt, D. & Schuhmann, L. Reconstitution of bacteriorhodopsin. FEBS Lett 44, 262-265 (1974). 14 Stoeckenius, W. & Bogomolni, R. A. Bacteriorhodopsin and related pigments of halobacteria. Annual review of biochemistry 51, 587-616 (1982). 15 Harrison, F. C. & Kennedy, M. E. The red discolouration of cured codfish. (FA Acland, printer, 1929). 16 Hampp, N. Bacteriorhodopsin as a photochromic retinal protein for optical memories. Chemical Reviews 100, 1755-1776 (2000). 17 Khorana, H. G. et al. Amino acid sequence of bacteriorhodopsin. Proceedings of the National Academy of Sciences 76, 5046-5050 (1979). 18 Dunn, R. et al. The bacteriorhodopsin gene. Proceedings of the National Academy of Sciences 78, 6744-6748 (1981). 19 Seehra, J. & Khorana, H. Bacteriorhodopsin precursor. Characterization and its integration into the purple membrane. Journal of Biological Chemistry 259, 4187-4193 (1984). 20 Balashov, S. P., Imasheva, E. S., Govindjee, R., Sheves, M. & Ebrey, T. G. Evidence that aspartate-85 has a higher pK (a) in all-trans than in 13-cisbacteriorhodopsin. Biophysical journal 71, 1973-1984 (1996). 21 Prokhorenko, V. I. et al. Coherent control of retinal isomerization in bacteriorhodopsin. Science 313, 1257-1261 (2006). 22 Birge, R. R. et al. Biomolecular electronics: protein-based associative processors and volumetric memories. The Journal of Physical Chemistry B 103, 10746-10766 (1999). 23 Onufriev, A., Smondyrev, A. & Bashford, D. Proton affinity changes driving unidirectional proton transport in the bacteriorhodopsin photocycle. Journal of molecular biology 332, 1183-1193 (2003). 24 Schobert, B., Cupp-Vickery, J., Hornak, V., Smith, S. O. & Lanyi, J. K. Crystallographic structure of the K intermediate of bacteriorhodopsin: conservation of free energy after photoisomerization of the retinal. Journal of molecular biology 321, 715-726 (2002). 25 Lanyi, J. K. & Schobert, B. Mechanism of proton transport in bacteriorhodopsin from crystallographic structures of the K, L, M1, M2, and M2' intermediates of the photocycle. Journal of molecular biology 328, 439 (2003). 26 Luecke, H. Atomic resolution structures of bacteriorhodopsin photocycle intermediates: the role of discrete water molecules in the function of this light-driven ion pump. Biochimica Et Biophysica Acta-Bioenergetics 1460, 133-156 (2000). 27 Luecke, H. et al. Coupling photoisomerization of retinal to directional transport in bacteriorhodopsin. Journal of molecular biology 300, 1237-1255 (2000). 28 Lanyi, J. K. Proton transfers in the bacteriorhodopsin photocycle. Biochimica et Biophysica Acta (BBA)-Bioenergetics 1757, 1012-1018 (2006). 29 Balashov, S. P., Imasheva, E. S., Govindjee, R. & Ebrey, T. G. Titration of aspartate-85 in bacteriorhodopsin: what it says about chromophore isomerization and proton release. Biophysical journal 70, 473-481 (1996). 30 Richter, H.-T., Brown, L. S., Needleman, R. & Lanyi, J. K. A linkage of the pKa's of Asp-85 and Glu-204 forms part of the reprotonation switch of bacteriorhodopsin. Biochemistry 35, 4054-4062 (1996). 31 Schobert, B., Brown, L. S. & Lanyi, J. K. Crystallographic structures of the M and N intermediates of bacteriorhodopsin: assembly of a hydrogen-bonded chain of water molecules between Asp-96 and the retinal Schiff base. Journal of molecular biology 330, 553-570 (2003). 32 Rouhani, S. et al. Crystal structure of the D85S mutant of bacteriorhodopsin: model of an O-like photocycle intermediate. Journal of Molecular Biology 313, 615-628 (2001). 33 Edman, K. et al. High-resolution X-ray structure of an early intermediate in the bacteriorhodopsin photocycle. Nature 401, 822-826 (1999). 34 Neutze, R. et al. Bacteriorhodopsin: a high-resolution structural view of vectorial proton transport. Biochimica et Biophysica Acta-Biomembranes 1565, 144-167 (2002). 35 Grzesiek, S. & Dencher, N. A. Time-course and stoichiometry of light-induced proton release and uptake during the photocycle of bacteriorhodopsin. FEBS letters 208, 337-342 (1986). 36 Gutman, M., Nachliel, E., Gershon, E., Giniger, R. & Pines, E. The pH jump: kinetic analysis and determination of the diffusion-controlled rate constants. Journal of the American Chemical Society 105, 2210-2216 (1983). 37 Gutman, M. Application of the laser-induced proton pulse for measuring the protonation rate constants of specific sites on proteins and membranes. Methods in Enzymology 127, 522-538 (1986). 38 Tamogami, J., Kikukawa, T., Miyauchi, S., Muneyuki, E. & Kamo, N. A tin oxide transparent electrode provides the means for rapid time-resolved pH measurements: application to photoinduced proton transfer of bacteriorhodopsin and proteorhodopsin. Photochemistry and photobiology 85, 578-589 (2009). 39 Kim, H. et al. Electrical, optical, and structural properties of indium–tin–oxide thin films for organic light-emitting devices. Journal of Applied Physics 86, 6451-6461 (1999). 40 Chu, L.-K., Yen, C.-W. & El-Sayed, M. A. Bacteriorhodopsin-based photo-electrochemical cell. Biosensors and Bioelectronics 26, 620-626 (2010). 41 Robertson, B. & Lukashev, E. P. Rapid pH change due to bacteriorhodopsin measured with a tin-oxide electrode. Biophysical journal 68, 1507-1517 (1995). 42 Wang, J.-P., Song, L., Yoo, S.-K. & El-Sayed, M. A. A comparison of the photoelectric current responses resulting from the proton pumping process of bacteriorhodopsin under pulsed and CW laser excitations. The Journal of Physical Chemistry B 101, 10599-10604 (1997). 43 Wang, J.-P., Yoo, S.-K., Song, L. & El-Sayed, M. A. Molecular mechanism of the differential photoelectric response of bacteriorhodopsin. The Journal of Physical Chemistry B 101, 3420-3423 (1997). 44 Saga, Y., Watanabe, T., Koyama, K. & Miyasaka, T. Mechanism of photocurrent generation from bacteriorhodopsin on gold electrodes. The Journal of Physical Chemistry B 103, 234-238 (1999). 45 Koyama, K. & Miyasaka, T. The proton uptake channel of bacteriorhodopsin as studied by a photoelectrochemical method. Bioelectrochemistry 53, 111-118 (2001). 46 Wu, J. et al. Efficient Approach to Determine the pKa of the Proton Release Complex in the Photocycle of Retinal Proteins. The Journal of Physical Chemistry B 113, 4482-4491 (2009). 47 Sass, H. J. et al. Structural alterations for proton translocation in the M state of wild-type bacteriorhodopsin. Nature 406, 649-653 (2000). 48 Hirai, T. & Subramaniam, S. Protein conformational changes in the bacteriorhodopsin photocycle: comparison of findings from electron and X-ray crystallographic analyses. PloS one 4, e5769 (2009). 49 Subramaniam, S. & Henderson, R. Molecular mechanism of vectorial proton translocation by bacteriorhodopsin. Nature 406, 653-657 (2000). 50 Andersson, M. et al. Structural dynamics of light-driven proton pumps. Structure 17, 1265-1275 (2009). 51 Nakatsuma, A. et al. Chimeric microbial rhodopsins containing the third cytoplasmic loop of bovine rhodopsin. Biophysical journal 100, 1874-1882 (2011). 52 Schätzler, B. et al. Subsecond proton-hole propagation in bacteriorhodopsin. Biophysical journal 84, 671-686 (2003). 53 Yang, C.-S., Sineshchekov, O., Spudich, E. N. & Spudich, J. L. The cytoplasmic membrane-proximal domain of the HtrII transducer interacts with the EF loop of photoactivated Natronomonas pharaonis sensory rhodopsin II. Journal of Biological Chemistry 279, 42970-42976 (2004). 54 Etzkorn, M. et al. Complex formation and light activation in membrane-embedded sensory rhodopsin II as seen by solid-state NMR spectroscopy. Structure 18, 293-300 (2010). 55 Scheerer, P. et al. Crystal structure of opsin in its G-protein-interacting conformation. Nature 455, 497-502 (2008). 56 Bryant, F. R. Construction of a recombinase-deficient mutant recA protein that retains single-stranded DNA-dependent ATPase activity. Journal of Biological Chemistry 263, 8716-8723 (1988). 57 Durfee, T. et al. The complete genome sequence of Escherichia coli DH10B: insights into the biology of a laboratory workhorse. Journal of bacteriology 190, 2597-2606 (2008). 58 Miroux, B. & Walker, J. E. Over-production of proteins in Escherichia coli: mutant hosts that allow synthesis of some membrane proteins and globular proteins at high levels. Journal of molecular biology 260, 289-298 (1996). 59 Ho, S. N., Hunt, H. D., Horton, R. M., Pullen, J. K. & Pease, L. R. Site-directed mutagenesis by overlap extension using the polymerase chain reaction. Gene 77, 51-59 (1989). 60 Vallejo, A. N., Pogulis, R. J. & Pease, L. R. PCR mutagenesis by overlap extension and gene SOE. Cold Spring Harbor Protocols 2008, pdb. prot4861 (2008). 61 傅喣媛. 表現 Haloarcula marismortui 之六個光感受體揭露其獨特的感光特性. 國立台灣大學碩士論文 (2008). 62 Fu, H.-Y., Chang, Y.-N., Jheng, M.-J. & Yang, C.-S. Ser262 determines the chloride-dependent colour tuning of a new halorhodopsin from Haloquadratum walsbyi. Bioscience Reports 32, 501-509 (2012). 63 沈宜中. HmBRI 突變設計蛋白質之光學特性與應用研究. 國立台灣大學碩士論文 (2012). 64 Dunn, R. et al. Structure-function studies on bacteriorhodopsin. I. Expression of the bacterio-opsin gene in Escherichia coli. Journal of Biological Chemistry 262, 9246-9254 (1987). 65 Mollaaghababa, R., Steinhoff, H.-J., Hubbell, W. L. & Khorana, H. G. Time-resolved site-directed spin-labeling studies of bacteriorhodopsin: loop-specific conformational changes in M. Biochemistry 39, 1120-1127 (2000). 66 Huang, K., Bayley, H., Liao, M.-J., London, E. & Khorana, H. Refolding of an integral membrane protein. Denaturation, renaturation, and reconstitution of intact bacteriorhodopsin and two proteolytic fragments. Journal of Biological Chemistry 256, 3802-3809 (1981). 67 Booth, P. J., Farooq, A. & Flitsch, S. L. Retinal binding during folding and assembly of the membrane protein bacteriorhodopsin. Biochemistry 35, 5902-5909 (1996). 68 Hoffmann, M. et al. Color tuning in rhodopsins: The mechanism for the spectral shift between bacteriorhodopsin and sensory rhodopsin II. Journal of the American Chemical Society 128, 10808-10818 (2006). 69 Brown, L. Proton transport mechanism of bacteriorhodopsin as revealed by site-specific mutagenesis and protein sequence variability. Biochemistry (Moscow) 66, 1249-1255 (2001). 70 Mogi, T., Stern, L. J., Marti, T., Chao, B. H. & Khorana, H. G. Aspartic acid substitutions affect proton translocation by bacteriorhodopsin. Proceedings of the National Academy of Sciences 85, 4148-4152 (1988). 71 Subramaniam, S., Greenhalgh, D. A., Rath, P., Rothschild, K. J. & Khorana, H. G. Replacement of leucine-93 by alanine or threonine slows down the decay of the N and O intermediates in the photocycle of bacteriorhodopsin: implications for proton uptake and 13-cis-retinal----all-trans-retinal reisomerization. Proceedings of the National Academy of Sciences 88, 6873-6877 (1991). 72 Brown, L. S., Needleman, R. & Lanyi, J. K. Conformational change of the EF interhelical loop in the M photointermediate of bacteriorhodopsin. Journal of molecular biology 317, 471-478 (2002). 73 Lanyi, J. K. Proton translocation mechanism and energetics in the light-driven pump bacteriorhodopsin. Biochimica et biophysica acta 1183, 241 (1993). 74 Heberle, J. Proton transfer reactions across bacteriorhodopsin and along the membrane. Biochimica et Biophysica Acta (BBA)-Bioenergetics 1458, 135-147 (2000). 75 Koyama, K., Miyasaka, T., Needleman, R. & Lanyi, J. K. Photoelectrochemical verification of proton‐releasing groups in bacteriorhodopsin. Photochemistry and photobiology 68, 400-406 (1998). | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61334 | - |
dc.description.abstract | 古細菌視紫紅質 (Bacteriorhodopsin;BR) 為存在嗜鹽古細菌 Halobacterium salinarum 膜上的蛋白質,其為一光驅動氫離子幫浦 (light-driven proton pump),照光後可將氫離子從細胞質打至細胞外。基態 BR 吸光後會依序轉變為各種中間過度態 (K、L、M、N 及 O),行使其功能後,最後回到基態,這個照光後引發的循環稱為光週期。在 BR 的光週期中,氫離子會在 BR 內部的殘基間傳遞,也會在 BR 內部的殘基和外界的水之間傳遞。在 L 中間態轉變為 M 中間態的過程中,BR 內部的氫離子釋放複合體 (proton releasing complex;PRC) 會先釋放一個氫離子至細胞外,之後,在 N 中間態轉變為 O 中間態的過程中,Asp 96 再從細胞內獲得一個氫離子。因此,一個能快速偵測水溶液 pH 值變化的裝置,對研究 BR 釋放及獲得氫離子的速率是必要的。本研究的第一部分先利用兩個已知為氫離子獲得速率較慢的突變蛋白 (E161C 及 R164C),確定了 BR-based 光電化學裝置在照光時的光電流衰減區 (decay of the light-on phase) 的衰減速率,以及停止照光後光電流恢復區 (recovery of the light-off phase) 的恢復速率,可以用來當作研究 wild type BR 及其突變蛋白獲得氫離子速率快慢的量化工具。而過去研究發現,BR 在行使功能的過程中 EF loop 會有顯著的構形變化,這表示 EF loop 很可能在 BR 行使功能時扮演了重要的角色。為了進一步研究 EF loop,本研究的第二部分將 EF loop 上的殘基單點突變為 Cys,並利用 BR-based 光電化學裝置和動態吸收光譜確定了 EF loop 上靠近 E-helix 及 F-helix 的 Cys 突變會顯著降低 BR 獲得氫離子的速率。這表示 EF loop 在 BR 獲得氫離子過程中扮演著重要的角色。 | zh_TW |
dc.description.abstract | Bacteriorhodopsin (BR) is the only protein in the purple membrane (PM) of Halobacteriun salinarum. BR is composed of seven transmembrane α-helices and contains one all-trans retinal chromophore which is covalently bound to the ϵ-amino group of the Lys 216 residue via a protonated Schiff base (PSB). Illumination of BR generates an electrochemical proton gradient across the PM by vectorial translocation of a proton from the cytoplasmic (CP) side to the extracellular (EC) side. Upon light absorption, the all-trans retinal isomerizes around the C13=C14 double-bond and reverts thermally back to the initial all-trans state, passing a series of intermediates named K, L, M, N, and O. The photocycle involves a proton transport process. During the L to M intermediate transition, a proton is released to the extracellular space from the proton releasing complex (PRC) which is composed of Arg 82, Glu 194, Glu 204, and internal H2O. During the N to O intermediate transition, the deprotonated Asp 96 uptake a proton from the cytoplasmic space. The transient pH change due to the light-induced proton release and uptake is usually studied by using a pH-sensitive dye whose absorption depends on pH. However, the restriction of the measurement is that the medium pH should be close to the pKa of the dye. Hence, a device is needed for the detection of the light-induced transient pH change in a wide pH range.
When illumination with a continuous wave (CW) laser pulse, a photocurrent is detected from the BR-based photoelectrochemical cell. At neutral pH, the positive photocurrent is observed when the CW light source is turned on, and the negative photocurrent is observed when the CW light source is turned off. The photocurrent is investigated for the transient proton concentration changes generated by BR. In previous study, it is found that the rates of the proton uptake decrease in the E161C and R164C mutant proteins using the pH-sensitive dye pyranine. In the first part of my work, I compare the photoresponse of the E161C and R164C mutations with the wild type BR. It is found that the decay rates of the light-on phase as well as the recovery rates of the light-off phase increase in the E161C and R164C mutations. These results suggest that the origin of the decay of the light-on phase as well as the recovery of the light-off phase are associated with the proton uptake of BR, and thus can be used for the investigation of the proton uptake rates of the wild type and mutant proteins. According to structural dynamic studies, significant conformational changes of the EF loop are observed during the photocycle of BR which means EF loop is likely critical in the function of BR. In the second part, I construct mutantions containing cysteine substitutions located in the EF loop of BR. Each of 10 residues, located in the EF loop of BR, was individually replaced by cystein, respectively. It is found that the rates of the proton uptake increase in the mutantions near the E-helix and F-helix. It means that EF loop play an important role during the proton uptake of BR. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T13:01:13Z (GMT). No. of bitstreams: 1 ntu-102-R00b22053-1.pdf: 5216772 bytes, checksum: fb4178355716f19c66a192157355dfd7 (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 目錄………………………………………………………………………………….…...i
圖目錄…………………………………………………………………………………...v 表目錄……………………………………………………………………………….…vii 摘要…………………………………………………………………………………....viii Abstract……………………………………………………………………………….....ix 第一章 緒論…………………………………………………………………………….1 第一節 微生物視紫紅質 (Microbial rhodopsin)………………………………....1 第二節 古細菌視紫紅質 (Bacteriorhodopsin; BR)………………………………3 2.1 古細菌視紫紅質的結構…………………………………………………3 2.2 古細菌視紫紅質的光週期 (Photocycle)………………………………..5 2.3 測量氫離子在蛋白質及外界水溶液間傳遞的重要性………………....9 第三節 對於 pH 值敏感的染劑可即時偵測水溶液 pH 值變化……………...9 第四節 以 BR 及 ITO 為材料建構光電池……………………....…………...11 第五節 EF loop 相關研究……………………………………………………....14 第六節 研究動機、設計與假設 (Hypothesis)…….………………………...…15 第二章 材料與方法………………………………………………………………...…17 第一節 實驗材料與藥品………………………………………………………...17 1.1 菌種……………………………………………………………………..17 1.2 質體…………………………………………………………………..…17 1.3 藥品……………………………………………………………………..17 第二節 實驗儀器與設備……………………………………………………...…18 2.1 核酸電泳設備…………………………………………………………..18 2.2 蛋白質電泳與轉印設備……………………………………………..…18 2.3 離心機…………………………………………………………………..19 2.4 光驅動氫離子幫浦活性量測用儀器…………………………………..19 2.5 動態吸收光譜量測用儀器……………………………………………..19 2.6 光電流量測用儀器……………………………………………………..19 2.7 其它……………………………………………………………………..20 第三節 實驗方法……………………………………………………………...…20 3.1 HsBR之突變株質體建構……...………………………..…………...…20 3.2 HsBR 及其突變蛋白之表現及純化…………………………………...25 3.2.1 蛋白質表現……………………………………………..…….........25 3.2.2 破菌、蛋白質粗萃取及膜分離……..………………..…………….25 3.2.3 回溶膜蛋白質…………............………………………………..….26 3.2.4 親和層析法……………………………………………..………….26 3.3蛋白質定性及定量基本分析…………………………...………………27 3.3.1 SDS - PAGE變性膠體電泳……………………...………………...27 3.3.2 CBR染色分析……………………………………………….……..27 3.3.3 西方墨點法…………………………………………………...……28 3.3.4 特徵吸收峰光譜鑑定…………...…………………………………28 3.4 氫離子幫浦活性測試……………………………………………..……28 3.5 動態吸收光譜量測…………………………………………………..…29 3.6 光電流量測…………………………………………………………..…29 第三章 結果與討論…...............................................................................................…30 第一節 BR-based 光電化學裝置在持續照光時光電流衰減區的衰減速率,以及停止照光後光電流恢復區的恢復速率,可以用來當作研究 BR 及其突變蛋白氫離子獲得速率的量化工具.............................................................................30 1.1 Halobacterium salinarum 的 BR 及其突變蛋白 E161C 及 R164C 可於大腸桿菌 C43 (DE3) 異源表現,且表現的 BR 及其突變蛋白皆具有光驅動氫離子幫浦的活性.................................................................................30 1.2 BR 及其突變蛋白 E161C 及 R164C 有正確的分子量.....................31 1.3 E161C 及 R164C 的最高特徵吸收峰和 WT 相近,和過去的研究結果一致.............................................................................................................32 1.4 E161C 獲得氫離子的速率較 WT 慢,和過去的研究結果一致..........33 1.5持續照光時的光電流衰減區的衰減速率,以及停止照光後光電流恢復區的恢復速率,可以用來當作研究 BR 及其突變蛋白獲得氫離子速率的量化工具.........................................................................................................34 第二節 EF loop 上靠近 E helix 及 F helix 的 Cys 突變會顯著降低 proton uptake 速率.............................................................................................................37 2.1 Halobacterium salinarum 的 BR 及其 EF loop 的 single Cys 突變蛋白可被大腸桿菌 C43 (DE3) 異源表現........................................................37 2.2 BR 及其 EF loop 的 single Cys 突變蛋白有正確的分子量..............40 2.3 EF loop 的 single Cys 突變蛋白的最高特徵吸收峰和 WT 相近......41 2.4分析光電流訊號發現,EF loop 上靠近 E helix 及 F helix 的 Cys 突變會顯著降低 BR 獲得氫離子的速率........................................................45 2.5 EF loop single Cys 突變蛋白 G155C、F156C、M163C 及 R164C 由 M 中間態回到基態所需的時間,明顯較 WT 來得長................................51 第四章 結論...................................................................................................................54 第五章 未來展望...........................................................................................................55 參考文獻.........................................................................................................................56 | |
dc.language.iso | zh-TW | |
dc.title | EF 環胺基酸單點突變改變 Halobacterium salinarum 古細菌視紫紅質之光電化學及動態吸光特性 | zh_TW |
dc.title | Single Residue Mutations at the EF Loop Affect the Properties of the Photocurrent and Dynamic Absorbance Spectrum of Halobacterium salinarum Bacteriorhodopsin | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 黃青真,張麗冠,梁博煌,李昆達 | |
dc.subject.keyword | 古細菌視紫紅質 獲得氫離子 BR-based 光電化學裝置 EF loop, | zh_TW |
dc.subject.keyword | Bacteriorhodopsin proton uptake BR-based photoelectrochemical cell EF loop, | en |
dc.relation.page | 62 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-08-07 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科技學系 | zh_TW |
顯示於系所單位: | 生化科技學系 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 5.09 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。