|
|||||
|
|
| 基于生物质谱技术的磷酰化修饰策略在多肽测序中的应用 | |
| 引用本文： | 张冬梅,刘红霞,毕开顺,陈晓辉,赵玉芬,蒋宇扬.基于生物质谱技术的磷酰化修饰策略在多肽测序中的应用[J].分析测试学报,2010,29(9). |
| 作者姓名： | 张冬梅 刘红霞 毕开顺 陈晓辉 赵玉芬 蒋宇扬 |
| 作者单位： | 1. 沈阳药科大学,药学院,辽宁,沈阳,110016;清华大学,深圳研究生院,广东省化学生物学重点实验室,广东,深圳,518055 2. 清华大学,深圳研究生院,广东省化学生物学重点实验室,广东,深圳,518055 3. 沈阳药科大学,药学院,辽宁,沈阳,110016 4. 厦门大学,化学化工学院,福建,厦门,316005 |
| 基金项目： | 国家自然科学基金资助项目 |
| 摘 要： | 该文建立了一种利用磷酰化修饰结合电喷雾质谱(ESI-Q-TOF)测定多肽氨基酸序列的有效方法。利用Atherton-Todd反应,以二丙基亚磷酰酯(DPP)为磷酰化试剂,应用生物质谱技术,对磷酰化修饰后的5种模型肽的磷酰化反应情况进行了系统研究,考察了磷酰化肽的二级质谱特征,并与未经磷酰化反应的肽的二级质谱特征对比。结果表明,经过磷酰化修饰后,肽的二级质谱中的a1离子信号强度明显增加,可以准确鉴定其N端氨基酸;b系列离子信息完整,信号强度增强,使得多肽C ID测序的谱图简单、清晰,有利于肽的氨基酸序列的测定;赖氨酸(K,128.10 u)和谷氨酰胺(Q,128.13 u)两种氨基酸质荷比相近,由于二者磷酰化修饰后的差异性,使其得到准确区分。经过5种已知氨基酸序列的模型肽的磷酰化后结合质谱技术进行氨基酸序列测定验证,结果表明该方法简单、快速、准确,提高了利用质谱技术进行多肽测序的准确度和灵敏度,可为蛋白质组学研究提供有效的技术手段。 |
| 关 键 词： | 二丙基亚磷酰酯 磷酰化修饰 从头测序 蛋白质组学 电喷雾质谱 |
Peptide Sequencing Based on Phosphorylation Strategy and Biological Mass Spectrometry Technology |
|
| ZHANG Dong-mei,LIU Hong-xia,BI Kai-shun,CHEN Xiao-hui,ZHAO Yu-fen,JIANG Yu-yang.Peptide Sequencing Based on Phosphorylation Strategy and Biological Mass Spectrometry Technology[J].Journal of Instrumental Analysis,2010,29(9). | |
| Authors: | ZHANG Dong-mei LIU Hong-xia BI Kai-shun CHEN Xiao-hui ZHAO Yu-fen JIANG Yu-yang |
| Affiliation: | ZHANG Dong-mei1,2,LIU Hong-xia2,BI Kai-shun1,CHEN Xiao-hui1,ZHAO Yu-fen3,JIANG Yu-yang2(1.School of Pharmacy,Shenyang Pharmaceutical University,Shenyang 110016,China,2.The Key Laboratory of Chemical Biology,Guangdong Province,Graduate School at Shenzhen,Tsinghua University,Shenzhen 518055,3.School of Chemistry and Chemical Engineering,Xiamen University,Xiamen 316005,China) |
| Abstract: | A simple and rapid method for determining peptide amino-acid sequence using ESI-Q-TOF-MS-based phosphorylation strategy was developed.Five model peptides were studied by ESI-Q-TOF-MS after modifying with dipropyl phosphonate(DPP) via the Atherton Todd reaction in the presence of tetrachloromethane and triethylamine.The characteristics of MS/MS spectra of phosphorylated peptide were studied and compared with those of non-phosphorylated peptides.Results showed that the signal intensity of a1 ion in the MS/MS ... |
| Keywords: | dipropyl phosphonate phosphorylation reaction de novo sequencing proteome research electrospray ionization mass spectrometry |
| 本文献已被 CNKI 万方数据 等数据库收录！ | |
