摘要
以糖蛋白核糖核酸酶B(RNase B)为研究对象,采用基质辅助激光解吸电离-串联飞行时间质谱(MALDI-TOF/TOF-MS)、傅里叶变换离子回旋共振质谱(FT-ICR-MS)的碰撞活化解离(CAD)和电子转移解离(ECD)多种质谱技术对RNase B的糖肽的完整信息进行解析。通过多种质谱相互验证,确定了RNase B的糖肽组分的氨基酸序列为SRNLTK、糖链结构为(Hex NAc)2(Hex)5-(Hex NAc)2(Hex)9及糖基化位点为第34位氨基酸残基。本方法为糖蛋白结构的全面解析提供了一种快速、有效、全面的研究思路。
The mass spectrometric analysis of both glycosylation site and glycan structure is an important and challenging topic in glycoprotein research. In this work,matrix-assisted laser desorption / ionization-time of flight / time of flight mass spectrometry（ MALDI-TOF / TOF-MS）,Fourier transform ion cyclotron resonance mass spectrometry（ FT-ICR-MS） combining with collision activation dissociation（ CAD） and electron capture dissociation（ ECD） MS technique were applied to analyze glycopeptide from ribonuclease B（ RNase B）digested by Lys-C and PNGase F. Through mutual validation by multiple mass spectrometric techniques,the amino acid sequence（ SRNLTK）, the glycosylation site（ AA34） and the glycan structure（（ Hex NAc）2（ Hex）5-（ Hex NAc）2（ Hex）9） of the glycopeptides were identified. Our method demonstrates to be a comprehensive,rapid and effective strategy in the glycoprotein analysis,and has a broad application prospect in the identification of unknown and complex glycoprotein.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2015年第4期564-569,共6页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金面上项目(No.21275134)资助~~
关键词
基质辅助激光解吸电离-串联飞行时间质谱
傅里叶变换离子回旋共振质谱
碰撞活化解离
电子捕获解离
核糖核酸酶B
Matrix-assisted laser desorption / ionization-time of flight / time of flight mass spectrometry
Fourier transform ion cyclotron resonance mass spectrometry
Collision activation dissociation
Electron capture dissociation
Ribonuclease B
