含有28个氨基酸的复杂多肽的串级质谱全序列分析研究.pdfVIP

2007 年第 65 卷 化 学 学 报 Vol. 65, 2007 第 3 期, 177～183 ACTA CHIMICA SINICA No. 3, 177～183 ·研究通讯· 含有 28 个氨基酸的复杂多肽的串级质谱全序列分析研究 贾韦韬 a,b 陆豪杰*,a,b 贠 栋 a,b 杨芃 原*,a,b (a 复旦大学化学系 上海 200433) (b 复旦大学生物医学研究院 上海 200032) 摘要 利用 MALDI-TOF/TOF MS 和 ESI-MS/MS 对一种含有多达 28 个氨基酸的复杂合成多肽成功进行了全序列测定. 通过调节激光强度、碰撞诱导解离(CID)能量等质谱参数以及依据不同序列分析软件, 获得了涵盖所有 b 型和 y 型碎片 离子的串级质谱图. 结果显示这种方法可以有效地解决 de novo 测序方法遇到的谱峰过于复杂导致运算死机等问题. 通 过讨论如何对含有超过 20 个氨基酸片断的多肽进行合格串级质谱实验, 为蛋白质组学肽段序列测定提供了新的方法 和思路. 关键词 串级质谱; 序列分析; 多肽 Sequence Analysis of a Complex Polypeptide Containing 28 Amino Acids by Tandem Mass Spectrometry JIA, Wei-Taoa,b LU, Hao-Jie*,a,b YUN, Donga,b YANG, Peng-Yuan*,a,b a ( Department of Chemistry , Fudan University, Shanghai 200433) (b Institutes of Biomedical Sciences, Fudan University, Shanghai 200032) Abstract One synthetic complex polypeptide composed of 28 amino acids was successfully sequenced by matrix-assisted laser desorption/ionization-time of flight/time of flight-mass spectrometry (MALDI-TOF/ TOF MS) and electrospray ionization-tandem mass spectrometry (ESI-MS/MS). Depending on the analysis of different sequencing software and the adjustment of the mass spectrometry parameters as well as the laser intensity, the collision induced dissociation (CID) energy and so on, we obtained the perfect tandem mass spectrum which covered all the b- and y-type fragment ions. Results indicated t