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標題: 裂解技術於環狀胜肽之質譜分析應用
Elucidation of CID and ETD based MS/MS Fragmentation for Analysis of Cyclic Peptides
作者: 吳家豪
Wu, Jia-Hau
Contributors: 賴建成
Chien-Chen Lai
分子生物學研究所
關鍵字: 環狀胜肽;質譜分析;碰撞引致裂解;電子轉移裂解
cyclic peptide;mass analysis;CID;ETD
日期: 2013
Issue Date: 2013-11-07 12:20:42 (UTC+8)
Publisher: 分子生物學研究所
摘要: 環狀胜肽 (cyclic peptide) 是由蛋白質或非蛋白質胺基酸藉由胜肽鍵所形成之環狀化合物,其種類繁多,並各自有不同功用,如可作為抗體、毒性物質、抗免疫物質、離子傳導調控物質、蛋白質鍵結抑制物或酵素抑制物。目前,環狀胜肽的應用相當多樣化,如使用於畜牧業作為其動物的促進生長藥物、農業上的天然農藥、天然界面活性劑等,甚至運用在高級化妝品、土壤復育、原油回收、幫助去除重金屬汙染等。對於環狀胜肽的相關研究,逐漸吸引學者的關注,本實驗的研究策略是以質譜分析技術為基礎並有別於前人研究以碰撞誘導裂解 (collision induced dissociation, CID) 分析環狀胜肽為主,嘗試運用其他較為新穎的裂解技術,包含電子轉移裂解 (electron transfer dissociation, ETD)、Electron-transfer collisionally activated dissociation (ETcaD) 及電子轉移結合碰撞引致裂解 (electron-transfer coupled with collision-induced dissociation, ET/CID) 進行環狀胜肽定性技術開發與探討。對於本身較不易帶有多價數之環狀胜肽,嘗試藉由添入金屬醋酸鹽溶液提升環狀胜肽於電灑法中電荷數,並觀察其是否可提升 ETD 的裂解效率,增加裂解資訊提高序列涵蓋率 (sequence coverage),使其擁有較高的可信度,建立一套環狀胜肽之質譜分析平台,協助未來相關菌種純化環狀胜肽之品質監測。
The cyclic peptide is one kind of cyclic compounds which combine of the protein and non-protein amino acids, many of kinds has more different functions, such as antibodies, toxic substances, anti-immune substances, ion transfer regulators, protein binding inhibitor or enzyme inhibitors. The application of the cyclic peptide is quite diverse, and gradually attracted the attention of scholars. The research strategy of this experiment is based on electron-spray ionization tandem mass spectrometry (ESI-MS/MS) combine with collision induced dissociation (CID) and electron transfer dissociation (ETD) to sequence cyclic peptides, to apply to the determination of cyclic peptides, Surfactin、Colistin、Polymyxin B、Iturin a and Bacitracin. Observations of different amino acid residues from CID and ETD experiments for the peptide were interpreted by their fragments, and try to join with metal acetate solution, that expect to enhance the number of electric charge for cyclic peptide in the electrospray ionization method, and observe whether increases the sequence coverage of ETD or not, to improve the qualitative degree the establishment of a cyclic peptide analysis platform, it might be the related bacteria purified cyclic peptide technology development and quantitative research.
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