摘要
蛋白质糖基化修饰位点的定位分析在糖基化蛋白质研究领域至关重要,本实验建立CID中性丢失引发的多级质谱对多肽糖基化位点分析的方法。以糖基化多肽为研究对象,采用ESI CID MS对Amadori糖基化多肽进行CID多级激发裂解,MS3裂解分别可以由3H2O、3H2O+HCHO的中性丢失引发,根据糖基化肽段的分析鉴定,均可完成对糖基化修饰位点的定位分析,但是在多级质谱分析中,3H2O中性丢失引发的糖基化肽段的鉴定更加精确,且在干热条件下,卵清蛋白中的K227相对K229更易发生糖基化修饰。综上分析,通过CID MS3多步激发质谱技术可以对一糖多肽进行糖基化定位分析。
Identification of the glycated sites in proteins plays an important role in the field of protein glycation. In the present study, we have established a CID neutral-loss-triggered MS3 method to analyze the glycated sites in proteins. ESI- CID-MS was used to stimulate multi-site cleavage in glycated Amadori peptides. MS^3 scans triggered by neutral losses of 3H2O and 3H2O ＋ HCHO produced similar results in terms of glycated peptide identification. However, neutral loss of 3H2O resulted in more accurate glycated peptide identification in multi-stage activation experiments. The results showed that the K227 was more easily glycated by D-glucose when compared with the K229 in ovalbumin under dry heating conditions. Overall, the multi-stage activation approach could identify the glycation site in peptides with one glycosylation site.
出处
《食品科学》
EI
CAS
CSCD
北大核心
2013年第23期47-50,共4页
Food Science
基金
国家"973"计划前期研究专项(2012CB126314)
关键词
糖基化肽
中性丢失
CID
多级质谱
glycated peptide
neutral-loss
collision-induced dissociation （CID）
multi-stage MS
作者简介
王辉（1982-），女，助理研究员，博士，研究方向为蛋白质的化学修饰及改性。E-mail：wanghui00072@aliyun．com
通信作者：涂宗财（1965-），男，教授，博士，研究方向为极端条件下食品蛋白质营养与安全。E-mail：tuzc_mail@aliyun．com
