頁籤選單縮合
題 名 | 以動力方法探索蛋白質構形之變化=Probing Conformational Changes of Protein by Kinetic Method |
---|---|
作 者 | 陳錦紋; 何永皓; | 書刊名 | 化學 |
卷 期 | 73:1 2015.03[民104.03] |
頁 次 | 頁13-22 |
專 輯 | 生物分析化學 |
分類號 | 341 |
關鍵詞 | 動力方法; 蛋白質構形; 離子遷移質譜法; Kinetic method; Protein conformation; Ion mobility mass spectrometry; |
語 文 | 中文(Chinese) |
中文摘要 | 蛋白質的構形變化影響其生物機能甚鉅,非自然構形之蛋白質常導致一些病變的發生,例如阿茲海默症、帕金森氏症、狂牛症等,研究蛋白質構形的方法有質譜、核磁共振、X-ray繞射法等,氫/氘交換和離子遷移法則為質譜法研究蛋白質構形時常用的策略。本實驗室則以動力方法來探索氣相蛋白質離子的構形變化,藉由蛋白質離子與參考物 EDTA所形成之質子化二聚體,經碰撞誘導解離後所產生之斷片離子相對強度,觀察不同電荷態蛋白質離子其質子競爭力的強弱。結果顯示 cytochrome c、lysozyme、ubiquitin、myoglobin等蛋白質其質子競爭力開始有明顯改變時的電荷態,恰巧與已知蛋白質離子由緊密摺疊轉為成部分緊密摺疊的電荷態一致,證實動力方法具有簡易、快速探索蛋白質構形的能力。 |
英文摘要 | Physiological properties of protein are affected by its conformation. Denatured conformation of protein could cause disease, such as Alzheimer’s disease, Parkinson’s disease, Creutzfeldt-Jakob disease. NMR, X-ray diffraction, and mass spectrometry are the techniques used for studying protein conformations. Hydrogen/deuterium exchange and ion mobility are typical methods of mass spectrometry to investigate protein conformation. We developed a new strategy to apply the kinetic method for probing protein conformations at different charge states. Proton-bound dimer produced from protein and EDTA was subject to collision-induced dissociation. Relative intensities of ion from two different dissociation paths can reflect the proton affinity of protein ion. Cytochrome c, lysozyme, ubiquitin, and myoglobin all showed a rapidly change of proton affinity at a special charge state. The charge state was inconsistent with that of the protein conformation transited from native to denatured form, which recorded by ion mobility method. Results indicated that the kinetic method has capability as a rapid and simple method for probing conformational changes of protein. |
本系統中英文摘要資訊取自各篇刊載內容。