請用此 Handle URI 來引用此文件: http://hdl.handle.net/11455/22018
標題: (1)建立以選擇電子轉移反應偵測模式的胜肽定量新方法(2)利用質譜儀鑑定水稻突變株M0047286榖粒外殼色素成分
(1)New Approaches for Quantitative Peptides Analysis by Selected Electron Transfer Reaction Monitoring (2)Characterization of Pigment in Rice Mutant M0047286 by Mass Spectrometry
作者: 魏碧瑩
Wei, Bi-Ying
關鍵字: mass spectrometry
電子轉移裂解
electron transfer dissociation
peptide quantitative
TNG67
pigment
serotonin
胜肽定量
台農67
色素
血清素
出版社: 分子生物學研究所
引用: (1) Aebersold, R., et al. (2001) Mass spectrometry in proteomics. Chemical Review, 101 : 269-295. Barnidge, D.R., et al. (2004) Absolute Quantification of the Model Biomarker Prostate-Specific Antigen in Serum by LC-MS/MS Using Protein Cleavage and Isotope Dilution Mass Spectrometry. Journal of Proteome Research, 3 : 644-652. Bunger, M.K., et al. (2008) Automated Proteomics of E. coli via Top-Down Electron-Transfer Dissociation Mass Spectrometry. Analysis Chemistry, 80 : 1459-1467. Chi, A., et al. (2007) Analysis of phosphorylation sites on proteins from Saccharomyces cerevisias by electron transfer dissociation (ETD) mass spectrometry. PNAS, 104 : 2193-2198. Coon, J.J., et al. (2005) Protein identification using sequential ion/ion reactions and tandem mass spectrometry. PNAS, 102 : 9463-9468. Domon B., et al. (2006) Mass Spectrometry and Protein Analysis. Science, 312 : 212-217. Fenn, J.B., et al. (1989) Electrospray ionization for mass spectrometry of large biomolecules. Science, 246 : 64-71. Fenselau, C., et al. (2007) A review of quantitative methods for proteomics studies. Journal of Chromatogrphy B, 855 : 14-20. Goshe, M.B., et al. (2003) Stable isotope-coded proteomic mass spectrometry. Current Opinion in Biotechnology, 14 : 101-109. Han, H., et al. (2008) Eletron Transfer Dissociation of iTRAQ Labeled Peptide Ions. Journal of Proteome Research, 7 : 3643-3648. Hillenkamp, F., et al. (1990) Mass spectrometry of peptides and proteins by matrix-assisted ultraviolet laser desorption/ionization. Methods Enzymol, 193 : 280-295. Hopfgartner, H., et al. (2005) New approaches for quantitative analysis in biological fluids using mass spectrometric detection. Trends in Analytical Chemistry, 24 : 583-589. Janecki, D.J., et al. (2007) A multiple reaction monitoring method for absolute quantification of the human liver alcohol dehydrogenase ADH1C1 isoenzyme. Analytical Biochemistry, 369 : 18-26. Kalli, A., et al. (2008) Comparison of the Electron Capture Dissociation Fragmentation Behavior of Doubly and Triply Protonated Peptides from Trypsin, Glu-C, and Chymotrypsin Digestion. Journal of Proteome Research, 7 : 2834-2844. Loo, J.A., et al. (1988) Collisional effects on the charge distribution of ions from large molecules, formed by electrospray-ionization mass spectrometry. Rapid Communications in Mass Spectrometry, 2 : 207-210. Madsen, J.A., et al. (2009) Simplifying Fragmentation Patterns of Multiply Charged Peptides by N-Terminal Derivatization and Electron Transfer Collision Activated Dissociation. Analysis Chemistry, 81 : 3645-3653. McLafferty, F.W., et al. (1981) Tandem Mass Spectrometry. Science, 214 : 280-287. McLafferty, F.W., et al. (2001) Electron Capture Dissociation of Gaseous Multiply Charged Ions by Fourier-Transform Ion Cyclotron Resonance. American Society for Mass Spectrometry, 12 : 245-249. Molina, H., et al. (2007) Global proteomic profiling of phosphopeptides using electron transfer dissociation tandem mass spectrometry. PNAS, 104 : 2199-2204. Molina, H., et al. (2008) Comprehensive Comparison of Collision Induced Dissociation and Electron Transfer Dissociation. Analysis Chemistry, 80 : 4825-4835. Phanstiel, D., et al. (2008) Peptide and Protein Quantification Using iTRAQ with Electron Transfer Dissociation. American Society for Mass Spectrometry, 19 : 1255-1262. Phanstiel, D., et al. (2009) Peptide Quantification Using 8-Plex Isobaric Tags and Electron Transfer Dissociation Tandem Mass Spectrometry. Analytical Chemistry, 81 : 1693-1698. Pitteri, S.J., et al. (2005) Electron-Transfer Ion/Ion Reactions of Doubly Protonated Peptides: Effect of Elevated Bath Gas Temperature. Analytical Chemistry, 77 : 5662-5669. Schlosser, A., et al. (2001) Analysis of protein phosphorylation by a combination of elastase digestion and neutral loss tandem mass spectrometry. Analytical Chemistry, 73 : 170-176. Shipkova, P., et al. (2008) Application of ion trap technology to liquid chromatography/mass spectrometry quantition of large peptides. Rapid Communications in Mass spectrometry, 22 : 1359-1366. Swaney, D.L., et al. (2007) Supplemental Activation Method for High-Efficiency Electron-Transfer Dissociation of Doubly Protonated Peptide Precursors. Analytical Chemistry, 79 : 477-485. Syka, J.E.P., et al. (2004) Peptide and protein sequence analysis by electron trandfer dissociation mass spectrometry. PNAS, 101 : 9528-9533. Tamvakopoulos, C. et al. (2007) Mass Spectrometry for the Quantification of Bioactive Peptides in Biological Fluids. Mass Spectrometry Reviews, 26 : 389-402. Xia, Y., et al. (2008) Activation of Intact Electron-Transfer Products of Polypeptides and Proteins in Cation Transmission Mode Ion/Ion Reactions. Analysis Chemistry, 80 : 1111-1117. Wang, H. et al. (2008) Accurate Localization and Relative Quantification of Arginine Methylation Using Nanoflow Liquid Chromatography Coupled to Electron Transfer Dissociation and Orbitrap Mass Spectrometry. American Society for Mass Spectrometry, 20 : 507-519. Watson, J.D., et al. (1990) The human genome project: past, present, and future. Science, 248 : 44-49. Wilm, M., et al. (1996) Parent ion scans of unseparated peptide mixtures. Analysis Chemistry, 68 : 527-533. Zhang, Q., et al. (2008) Improved Methods for the Enrichment and Analysis of Glycated Peptides. Analysis Chemistry, 80 : 9822-9829. Zubarev, R.A., et al. (1998) Electron Capture Dissociation of Multiply Charged Protein Cations. A Nonergodic Process. Journal of the American Chemical Society, 120 : 3265-3266. (2) Cao, J., et al. (2006) Rapid method for accurate analysis of melatonin, serotonin and auxin in plant samples using liquid chromatography-tandem mass spectrometry. Journal of Chromatography A, 1134 : 333-337. Domingues, M.R.M., et al. (2003) Identification of Oxidation Products and Free Radicals of Tryptophan by Mass spectrometry. American Society for Mass Spectrometry, 14 : 406-416. Dubouzet, J.G., et al. (2007) Integrated metabolomic and transcriptomic analyses of high-tryptophan rice expressing a mutant anthranilate synthase alpha subunit. Journal of Experimental Botany, 58 : 3309-3321. Facchini, P.J., et al. (2000) Plant aromatic L-amino acid decarboxylase: evolution, biochemistry, regulation, and metabolic engineering applications. Phytochemistry, 54 : 121-138. Favretto, D., et al. (1998) The Role of Peroxidase in the Oligomerization of 5-Hydroxytryptamine Investigated by Matrix-assisted Laser Desorption/Ionization Mass Spectrometry. Rapid Communication in Mass Spectrometry, 12 : 193-197. Griffin, T.J., et al. (2007) iTRAQ Reagent-Based Quantitative Proteomic Analysis on a Linear Ion Trap Mass Spectrometer. Journal of Proteome Research, 6 : 4200-4209. Huether, G., et al. (1997) Serotonin Acts as Radical Scavenger and Is Oxidized to a Dimer During the Respiratory Burst of Activated Microglia. Journal of Neurochemistry, 69 : 2096-2101. International Rice Genome Sequencing Project (2005) The map-based sequence of the rice genome. Nature, 436 : 793-800. Ishihara, A., et al. (2008) The tryptophan pathway is involved in the defense responses of rice against pathogenic infection via serotonin production. The Plant Journal, 54 : 481-495. Jang, S.M., et al. (2004) Production of Coumaroylserotonin and Feruloylserotonin in Transgenic Rice Expressing Pepper Hydroxycinnamoyl-Coenzyme A: Serotonin N-(Hydroxycinnamoyl)transferase. Plant Physiology, 135 : 346-356. Jeong, D.H., et al. (2002) T-DNA insertional metagenesis for activation tagging in rice. Plant Physiology, 130 : 1636-1644. Jeong, D.H., et al. (2006) Generation of a flanking sequence-tag database for activation-tagging lines in japonica rice. The Plant Journal, 45 : 123-132. Jones, C.E., et al. (2007) Copper induced oxidation of serotonin: analysis of products and toxicity. Journal of Neurochemistry, 102 : 1035-1043. Kang, S., et al. (2006) Functional Analysis of the Amine Substrate Specificity Domain of Pepper Tyramine and Serotonin N-Hydroxycinnamoyltransferases. Plant Physiology, 140 : 704-715. Kang, S., et al. (2007) Characterization of rice tryptophan decarboxylases and their direct involvement in serotonin biosynthesis in transgenic rice. Planta, 227 : 263-272. Krysan, P.J., et al. (1999) T-DNA as an Insertional Mutagen in Arabidopsis. The Plant Cell, 11 : 2283-2290. Kurata, N., et al. (2005) Rice mutants and genes related to organ development, morphogenesis and physiological traits. Plant and Cell Physiology, 46 : 48-62. Lee, K., et al. (2008) Endosperm-Specific Expression of Serotonin N-Hydroxycinnamoyltransferase in Rice. Plant Foods for Human Nutrition, 63 : 53-57. Loo, J.A., et al. (1988) Collisional effects on the charge distribution of ions from large molecules, formed by electrospray-ionization mass spectrometry. Rapid Communications in Mass Spectrometry, 2 : 207-210. Minki, D., et al. (2005) RNA silencing of single and multiple members in a gene family of rice. Plant Physiology, 138 : 1903-1913. Rizzi, A., et al. (2006) An investigation on the possibile role of melatonin in melanogensis. Journal of Mass Spectrometry, 41 : 517-526. Semak, I., et al. (2004) Serotonin metabolism in rat skin : characterization by liquid chromatography-mass spectrometry. Archive of Biochemistry and Biophysics, 421 : 61-66. Smaniotto, A., et al. (2006) A mass spectrometric investigation on the possible role of tryptophan and 7-hydroxytryptophan in melannogensis. Journal of Mass Spectrometry, 41 : 921-930. Tanaka, E., et al. (2003) Phenylpropanoid amides of serotonin accumulate in witches' broom diseased bamboo. Phytochemistry, 64 : 965-969. Zhang, W.J., et al. (2009) The effect of ultraviolet radiation on the accumulation of medicinal compounds in plants. Fitoterapia, 80 : 207-218.
摘要: (1) 液相層析串聯式質譜儀 (LC/MS/MS) 搭配選擇反應偵測模式 (Selective Reaction Monitoring,SRM) 是質譜分析中常用於胜肽定量的一套策略,此方法能專一針對特定的胜肽片段進行監測。SRM 是先選擇特定質荷比的母離子進行碰撞引致裂解 (Collision-induced dissociation,CID) 後,偵測特定質荷比的子離子片段。低能碰撞引致裂解是目前最常使用的裂解方法,在進行胜肽碎裂的反應會打斷胜肽鍵,利於進行胜肽的序列分析。但胜肽利用碰撞引致裂解方法碎裂之後,母離子會平均碎裂成子離子,因而大幅減低子離子強度。上述的情形容易影響 SRM 的靈敏度。電子轉移裂解 (Electron transfer dissociation,ETD) 為新發展的裂解方法,可補足碰撞引致裂解的缺點,保留蛋白轉譯後修飾的官能基,目前已成為研究蛋白轉譯後修飾的有利工具。然而電子轉移裂解應用在正二價胜肽離子時,經電子轉移後之胜肽離子裂解效率差。在此將電子轉移裂解針對正二價胜肽離子會產生電子轉移而無法有效裂解的缺點加以應用,以發展一套利用電子轉移裂解搭配 SRM 來做為胜肽定量的新方法,命名為選擇電子轉移反應偵測模式 (Selected electron transfer reaction monitoring,SETRM),即選擇特定質荷比的母離子,經電子轉移後偵測電荷還原離子。經實驗證實,針對正二價或正三價的母離子進行定量時,利用 SETRM 的方法能產生比 SRM 更強的離子訊號,因為一般以胰蛋白酶進行水解的胜肽於 LC/MS 分析中,主要為正二價或正三價的母離子為主,因此更適用此套定量方法。其中以 [Glu1]-Fibrinopeptide B Human 標準品測試 SRM 與 SETRM 的定量效果,發現 SETRM 與 SRM 都具有 3 個維度 (order) 的線性範圍,此外因為 SETRM 的定量方法能得到比 SRM 更高的離子訊號,因此期望利用此策略提升質譜儀在胜肽定量上的靈敏度,做為碰撞引致裂解定量時另一互補的方法,提升電子轉移裂解在質譜分析的應用效率。 (2) M0047286為 T-DNA 插入水稻TNG 67所產生的突變株,此突變株具有深褐色穀粒及外殼的外表型。前人研究證實 M0047286 其 286-10 與286-14 基因受到 T-DNA 活化。以酵素活性分析,證實 286-10 與 286-14 皆具有tryptophan decarboxylase (TDC) 的活性,TDC 可將tryptophan 轉換成為 tryptamine。利用基因轉殖技術,獲得能大量表現 286-10 與 286-14 的轉殖株 (Ubi::286-10 與 Ubi::286-14),同樣會產生含有褐色穀粒及外殼的外表型。在本研究中以質譜儀鑑定出在 M0047286 突變株與轉殖株中 serotonin 的含量皆比野生株 (TNG 67) 高。利用試管內進行 serotonin 氧化測試證實,以過氧化氫反應後會產生黃色的色素成分,而經 UV 刺激後則會產生褐色的色素。以質譜分析照射 UV 後變色的 serotonin 標準品,可偵測到 serotonin 形成二聚體的結構。利用液相層析串聯質譜儀分析證實,二聚體的結構在 M0047286 突變株、Ubi::286-10 與 Ubi::286-14的水稻外殼中含量皆高於野生株 (TNG 67)。目前的研究結果無法確定二聚體的結構,但由結果顯示二聚體的含量似乎與水稻外殼的色素變化相關,因此推測 serotonin 二聚體極有可能改變水稻外殼色素。
(1) Liquid chromatography coupled to tandem mass spectrometry (LC/MS/MS) approach with selective reaction monitoring (SRM) allowed for the specific and sensitive quantitation of peptides. SRM is achieved via MS/MS utilizing collision-induced dissociation (CID) while monitoring unique precursor to product ion transitions. Low-energy CID tandem mass spectrometry has been, by far, the most common method used to dissociate peptide ions for sequence analysis. However, collisional scattering of product ions in CID results in decrease in intensity of the primary product ion. The lower intensity of the targeted product ion can lead to a reduction in the sensitivity of a quantitative method with SRM. Electron transfer dissociation (ETD) is a prevailing fragmentation method that can be complementary to CID and its utility in sequencing peptides containing post-translational modification. During the ETD reaction, there is a significantly shift toward nondissociative electron transfer as function of decreasing precursor ion charge for doubly charged peptides. In this study, we have developed a method utilizing ETD while monitoring unique precursor to charge reduced ion called selective electron transfer reaction monitoring (SETRM). For +2 and +3 precursor ion, more intense targeted ions for SETRM than SRM that is more suitable to quantify for trypsin digestion peptides. For [Glu1]-Fibrinopeptide B Human, 3 order linearity and excellent accuracy are observed in both SRM and SETRM. Moreover, SETRM provided more signal intensity, expecting to detect lower Limit of detection (LOD) due to a less signal loss compared to SRM analysis. Therefore, a new peptide qualitative and quantitative approach utilizing ETD technique as proteomic tool is provided here. (2) M0047286 is a rice TNG67 T-DNA insertion mutant showed dark-brown color in leaves, grains and hulls. Genes 286-10 and 286-14 flanked by the T-DNA insertion site is activated in the previous studies. Enzymatic assay using specific substracte by HPLC revealed that 286-10 and 286-14 had tryptophan decarboxylase (TDC) activity, which catalyzes the conversion of tryptophan into tryptamine. Using transgenic approach, transgenic rice Ubi::286-10 and Ubi::286-14 also showed dark-brown hulls. This study LC/MS/MS demonstrated that M0047286 and transgenic rice (Ubi::286-10 and Ubi::286-14) had serotonin compound higher than TNG 67. In vitro oxidation of serotonin by H2O2 and UV radiation results in the eventual formation of yellow and brown pigments, respectively. Mass spectrometry provided evidence for the dimeric serotonin species from UV radiation studies. The serotonin dimer compounds were also detected in M0047286 mutant plant and transgenic rice in LC/MS/MS studies. Some isoforms were detected in the serotonin dimeric sprcies The structure of these isoforms need to be elucidated in the future. In conclusion, the dimeric species of serotonin is strongly related to pigment on rice.
URI: http://hdl.handle.net/11455/22018
其他識別: U0005-1707200914555900
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1707200914555900
顯示於類別:分子生物學研究所

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