食品科学 ›› 2013, Vol. 34 ›› Issue (23): 47-50.doi: 10.7506/spkx1002-6630-201323011

• 基础研究 • 上一篇    下一篇

利用CID MS3质谱的中性丢失分析多肽糖基化位点

王 辉1,涂宗财1,2,*,刘光宪3,沙小梅1,肖 辉4   

  1. 1.南昌大学 食品科学与技术国家重点实验室,江西 南昌 330047;2.江西师范大学生命科学学院,江西 南昌 330022;
    3.江西省农业科学院,江西 南昌 330200;4. 叶史瓦大学美国爱因斯坦医学院,纽约 10461,美国
  • 收稿日期:2013-04-09 修回日期:2013-11-13 出版日期:2013-12-15 发布日期:2014-01-03
  • 通讯作者: 涂宗财 E-mail:tuzc_mail@yahoo.com.cn
  • 基金资助:

    国家“973”计划前期研究专项(2012CB126314)

Analysis of Glycated Peptides by Neutral-loss-triggered CID MS3 Spectrometry

WANG Hui1,TU Zong-cai1,2,*,LIU Guang-xian3,SHA Xiao-mei1,XIAO Hui4   

  1. 1. State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047, China;
    2. College of Life Sciences, Jiangxi Normal University, Nanchang 330022, China;3. Jiangxi Academy of Agricultural Sciences,
    Nanchang 330200, China;4. Albert Einstein College of Medicine, Yeshiva University, New York 10461, USA
  • Received:2013-04-09 Revised:2013-11-13 Online:2013-12-15 Published:2014-01-03
  • Contact: tuzongcai zong-caiTU E-mail:tuzc_mail@yahoo.com.cn

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。

关键词: 糖基化肽, 中性丢失, CID, 多级质谱

Abstract:

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. ESICID-
MS was used to stimulate multi-site cleavage in glycated Amadori peptides. MS3 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.

Key words: glycated peptide, neutral-loss, collision-induced dissociation (CID), multi-stage MS

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