Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/92241
標題: Simultaneous Qualitative and Quantitative Analysis of Cyclic Peptides by CID and ETD Based MS/MS Fragmentation
建立以多種質譜裂解技術於環狀胜肽之同時定性與定量方法
作者: 阮馨平
Sin-Ping Ruan
關鍵字: 液相層析;質譜分析;環狀胜肽;電子轉儀裂解;LC-MS/MS;Cyclic-peptide;ETD
引用: 1. Wan E.C., Ho C., Sin D.W., Wong Y.C. Detection of residual bacitracin A, colistin A, and colistin B in milk and animal tissues by liquid chromatography tandem mass spectrometry. Anal Bioanal. Chem. 2006, 385, 181–188. 2. Kim P.I., Ryu J., Kim Y.H., Chi Y.T. Production of biosurfactant lipopeptides Iturin A, fengycin and surfactin A from Bacillus subtilis CMB32 for control of Colletotrichum gloeosporioides. J. Microbiol Biotechnol. 2010, 20, 138–45. 3. Yang Y.H., Fu S.G., Peng H., Shen A.D., Yue S.J., Go Y.F., Yuan L., Jiang Z.F. Abuse of antibiotics in China and its potential interference in determining the etiology of pediatric bacterial diseases. Pediatr Infect Dis J. 1993, 12, 986–988. 4. 吳家豪,2013,裂解技術於環狀胜肽之質譜分析應用,中興大學分子生物學研究所碩士論文。 5. Jones A.W., Cooper H.J. Dissociation techniques in mass spectrometry-based proteomics. Analyst. 2011, 136, 3419–3429. 6. Gause G.F, Brazhnikov A.M.G. Gramicidin S and its use in the Treatment of Infected Wounds. Nature. 1944 , 154, 703–709. 7. Craik D.J. Circling the enemy: cyclic proteins in plant defence. Trends Plant Sci. 2009, 14, 328–335. 8. Keymanesh K, Soltani S, Sardari S. Application of antimicrobial peptides in agriculture and food industry. World J. Microbiol. Biotechnol. 2009, 25, 933–944. 9. Matyus E, Kandt C, Tieleman DP. Computer simulation of antimicrobial peptides. Curr Med Chem. 2007, 14, 2789–2798. 10. Hathout Y., Ho Y.P., Ryzhov V., Demirev P., Fenselau C. Kurstakins: a new class of lipopeptides isolated from Bacillus thuringiensis. J Nat Prod. 2000, 63, 1492–1496. 11. Ongena M., Jourdan E., Adam A., Paquot M., Brans A., Joris B., Arpigny J.L., Thonart P. Surfactin and fengycin lipopeptides of Bacillus subtilis as elicitors of induced systemic resistance in plants. Environ Microbiol. 2007, 4, 1084–1090. 12. Meena K.R., Kanwar S.S. Lipopeptides as the antifungal and antibacterial agents: applications in food safety and therapeutics. Biomed Research International. 2015, 2015, 1–9. 13. Kim M.S.,LIim J.H., PARK B.K. Effect of Surfactin on growth performance of weaning piglets in combination with Bacillus subtilis BC1212. Journal of Veterinary Clinics. 2009, 2, 117–122. 14. Edman P. Method for the determination of the amino acid sequence in peptides. Acta Chem. Scand. 1950, 22, 283–293. 15. Volpon L., Besson F., Lancelin J.M. NMR structure of active and inactive forms of the sterol-dependent antifungal antibiotic bacillomycin L. Eur J Biochem. 1999, 264, 200–210. 16. Mohimani H., Yang Y.L., Liu W.T., Hsieh P.W., Dorrestein P.C., Pevzner P.A. Sequencing cyclic peptides by multistage mass spectrometry. Proteomics. 2011, 11, 3642–3650. 17. Gross M.L., McCrery D., Crow F., Tomer K.B., Pope M.R., Cuifetti L.M., Knoch H.W., Daly J.M., Dunkle L.D. The structure of the toxin from Helminthosporium carbonum. Tetrahedron. Lett. 1982, 51, 5381–5384. 18. Ngoka L.C., Gross M.L. Multistep tandem mass spectrometry for sequencing cyclic peptides in an ion-trap mass spectrometer. J Am Soc Mass Spectrom. 1999, 10, 732–746. 19. Govaerts C., Rozenski J., Orwa J., Roets E., Schepdael A.V., Hoogmartens J. Mass spectrometric fragmentation of cyclic peptides belonging to the polymyxin and colistin antibiotics studied by ion trap and quadrupole/orthogonal-acceleration time-of-flight technology. Rapid Commun Mass Spectrom. 2002, 9, 823–833. 20. Govaerts C., Li C., Orwa J., Schepdael A.V., Adams E., Roets E., Hoogmartens J. Sequencing of bacitracin A and related minor components by liquid chromatography/electrospray ionization ion trap tandem mass spectrometry. Rapid Commun Mass Spectrom. 2003, 17, 1366–1379. 21. Syka J.E.P., CoonJ.J, Schroeder M.J., Shabanowitz J., Hunt D.F. Peptide and protein sequence analysis by electron transfer dissociation mass spectrometry. PNAS. 2004, 101, 9528–9533. 22. Duan X., Engler F.A., Qu J. Electron transfer dissociation coupled to an Orbitrap analyzer may promise a straightforward and accurate sequencing of disulfide-bridged cyclic peptides: a case study. J Mass Spectrom. 2010, 45, 1477–1482. 23. Guan F., Uboh C.E., Soma L.R., Rudy J. Sequence elucidation of an unknown cyclic peptide of high doping potential by ETD and CID tandem mass spectrometry. J Am Soc Mass Spectrom. 2011, 22, 718–730. 24. Seidler J., Zinn N., Boehm M.E., Lehmann W.D. De novo sequencing of peptides by MS/MS. Proteomics. 2010, 4, 634–649. 25. Sin D.W.M., Ho C., Wong Y.C. Analysis of major components of residual bacitracin and colistin in food samples by liquid chromatography tandem mass spectrometry. Analytica Chimica Acta. 2005, 535, 23–31. 26. Ma Z., Wang J., Gerber J.P., Milne R.W. Determination of colistin in human plasma, urine and other biological samples using LC-MS/MS. J Chromatogr B Analyt Technol Biomed Life Sci. 2008, 862, 205–212. 27. Kaufmann A., Widmer M. Quantitative analysis of polypeptide antibiotic residues in a variety of food matrices by liquid chromatography coupled to tandem mass spectrometry. Anal Chim Acta. 2013, 797, 81–88. 28. Shields S.J., Bluhm B.K., Russell D.H. Fragmentation chemistry of [M + Cu]+ peptide ions containing an N-terminal arginine. J Am Soc Mass Spectrom. 2000, 11, 626–638. 29. Williams S.M., Brodbelt J.S. MS(n) characterization of protonated cyclic peptides and metal complexes. J Am Soc Mass Spectrom. 2004, 7, 1039–1054. 30. Fung Y.M., Liu H., Chan T.W. Electron capture dissociation of peptides metalated with alkaline-earth metal ions. J Am Soc Mass Spectrom. 2006, 6, 757–771. 31. Dong J., Vachet R.W. Coordination sphere tuning of the electron transfer dissociation behavior of Cu(II)-peptide complexes. J Am Soc Mass Spectrom. 2012, 2, 321–329. 32. Smith R.D., Barinaga C.J., Udseth H.R. Improved electrospray ionization interface for capillary zone electrophoresis-mass spectrometry. Anal Chem. 1988, 60, 1948–1952. 33. Charles L., Laure F., Raharivelomanana P., Bianchini J.P. Sheath liquid interface for the coupling of normal-phase liquid chromatography with electrospray mass spectrometry and its application to the analysis of neoflavonoids. J Mass Spectrom. 2005, 40, 75–82. 34. Girod M., Delaurent C., Charles L. Analysis of amitrole by normal-phase liquid chromatography and tandem mass spectrometry using a sheath liquid electrospray interface. Rapid Commun Mass Spectrom. 2006, 20, 892–896. 35. Ngoka L.C., Gross M.L. A nomenclature system for labeling cyclic peptide fragments. J Am Soc Mass Spectrom. 1999, 10, 360–363. 36. Zubarev R.A., Kelleher N.L., McLafferty F.W. Electron capture dissociation of multiply charged protein cations. A nonergodic process. J. Am. Chem. Soc. 1998, 13, 3265–3266. 37. Bakhtiar R., Guan Z. Electron capture dissociation mass spectrometry in characterization of peptides and proteins. Biotechnol Lett. 2006, 14, 1047–1059. 38. Jensen C.S., Wyer J.A., Houm?ller J., Hvelplund P., Nielsen S.B. Electron-capture induced dissociation of doubly charged dipeptides: on the neutral losses and N-Cα bond cleavages. Phys Chem Chem Phys. 2011, 41, 18373–18378. 39. Kim M.S., Pandey A. Electron transfer dissociation mass spectrometry in proteomics. Proteomics. 2012, 4-5, 530–542. 40. Wiesner J., Premsler T., Sickmann A. Application of electron transfer dissociation (ETD) for the analysis of posttranslational modifications. Proteomics. 2008, 21, 4466–4483. 41. Xia Y., Gunawardena H.P., Erickson D.E., McLuckey S.A. Effects of cation charge-site identity and position on electron-transfer dissociation of polypeptide cations. J Am Chem Soc. 2007, 40, 12232–12243. 42. Mikesh L.M., Ueberheide B., Chi A., Coon J.J., Syka J.E., Shabanowitz J., Hunt D.F. The utility of ETD mass spectrometry in proteomic analysis. Biochim Biophys Acta. 2006 , 12, 1811–1822. 43. Swaney D.L., McAlister G.C., Wirtala M., Schwartz J.C., Syka J.E., Coon J.J. Supplemental activation method for high-efficiency electron-transfer dissociation of doubly protonated peptide precursors. Anal Chem. 2007, 2, 477–485. 44. Chan W.Y., Chan T.W., O'Connor P.B. Electron transfer dissociation with supplemental activation to differentiate aspartic and isoaspartic residues in doubly charged peptide cations. J Am Soc Mass Spectrom. 2010, 6, 1012–1015. 45. Liu C.W., Lai C.C. Effects of electron-transfer coupled with collision-induced dissociation (ET/CID) on doubly charged peptides and phosphopeptides. J Am Soc Mass Spectrom. 2011, 1, 57–66. 46. McAlister G.C, Phanstiel D., Good D.M., Berggren W.T., Coon J.J. Implementation of electron-transfer dissociation on a hybrid linear ion trap-orbitrap mass spectrometer. Anal Chem. 2007, 10, 3525–3534.
摘要: 
The cyclic peptide is one kind of cyclic compounds which combine of the protein and non-protein amino acids, and the carboxyl function at the C-terminus of a peptide forms a peptide bond with the N-terminal amine group a cyclic peptide is formed. A wide variety of cyclic peptides, and all has different biological functions, such as antibodies, anti-bacterial, anti-cancer, anti-immune substances, toxic substances, ion transfer regulators or protein binding inhibitor. The application of the cyclic peptide is quite diverse, for example: animal feed additive, biopesticide, surfactants, cosmetic or human drug. In the past few years, because the overuse of antibiotics, the cyclic peptides from microorganism are gradually attracted the attention of scholars. The research strategy of this experiment is based on mass analysis, using liquid chromatography tandem mass spectrometry (LC-MS/MS) combine with selected reaction monitoring (SRM) to quantitative of five cyclic peptides antibiotics (polymyxin b, colistin a, colistin b, surfactin, iturin a) in a variety of matrices (bacterial, liver and milk), and combine with electron-transfer coupled with collision-induced dissociation (ET/CID) to sequence cyclic peptides, trying to join with metal chloride solution by sheath liquid, that expect to enhance the number of electric charge for cyclic peptides in the electrospray ionization method, and observe whether increases the sequence coverage of ETD, to establishment of a cyclic peptide containing such compounds ET/CID and SRM analysis platform.

環狀胜肽 (cyclic peptide) 是一種較線性多肽更為穩定且具有生理功能和醫藥價值的環狀多肽,由蛋白質或非蛋白質胺基酸藉由酯鍵、胜肽鍵、雙硫鍵所形成之環狀化合物,其廣泛分布、種類繁多且具不同功用,在鎮靜、抗菌、抗腫瘤、免疫抑制、抗酵素水解、抗化學降解、離子載體系統或蛋白質鍵結抑制物等方面展現出豐富多樣的生物活性。近年來,因化學性抗菌藥劑的長期濫用,使微生物來源的環狀胜肽變成潛在抗生素理想替代品之一,成為近期研究趨勢。目前,環狀胜肽的應用漸趨多樣化,如使用於畜牧業作為動物的飼料添加物、農業上的天然農藥、天然界面活性劑,甚至運用在高級化妝品、人類的醫學用藥等。於此以質譜分析技術為基礎,利用液相層析串聯式質譜儀 (liquid chromatography tandem mass spectrometry, LC-MS/MS) 搭配新穎裂解技術電子轉移碰撞引致裂解 (ET/CID) 達到良好定性結果,並結合選擇離子反應偵測模式 (selected reaction monitoring, SRM) 針對環狀胜肽作定量分析,在 ET/CID 定性方面,本身帶有單價數之環狀胜肽母離子,藉由鞘流溶液 (sheath liquid ) 方式添加金屬離子來提升其環狀胜肽之電荷數,提高對電子式裂解之分析效率。在 SRM 定量方面,偵測極限 (limit of detection, LOD) 在 S/N ratio > 3 為 0.41 nM ,定量極限 (limit of quantification, LOQ) 在 S/N ratio > 10 為 1.3 nM ,且變異係數 (CV%) 介於 3 ~ 11 % ;利用此分析平台可檢測出菌液中環狀胜肽含量,且線性範圍 r2 > 0.95 ;此外,針對市售牛奶與豬肝組織,皆未檢測出環狀胜肽,其線性範圍 r2 > 0.95。在本研究結果中,證實以鞘流溶液方式添加金屬離子可增加電荷數,應用於 ET/CID 此電子式裂解模式上,增加裂解資訊提高序列涵蓋率 (sequence coverage),使定性結果擁有較高的可信度,並同時進行定量,建立一個可針對含有環狀胜肽這類化合物的 ET/CID 定性與 SRM 定量分析平台。
URI: http://hdl.handle.net/11455/92241
Rights: 同意授權瀏覽/列印電子全文服務,2018-08-26起公開。
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