Label-free relative quantification of alpha-2-macroglobulin site-specific core-fucosylation in pancreatic cancer by LC-MS/MS

被引：28

作者：

Lin, Zhenxin ^{[1
]}

Yin, Haidi ^{[2
]}

Lo, Andy ^{[2
]}

Ruffin, Mack T. ^{[3
]}

Anderson, Michelle A. ^{[4
]}

Simeone, Diane M. ^{[2
]}

Lubman, David M. ^{[1
,2
]}

机构：

[1] Univ Michigan, Dept Chem, Ann Arbor, MI 48109 USA

[2] Univ Michigan, Med Ctr, Dept Surg, Ann Arbor, MI 48109 USA

[3] Univ Michigan, Med Ctr, Dept Family Med, Ann Arbor, MI 48109 USA

[4] Univ Michigan, Med Ctr, Dept Internal Med, Div Gastroenterol, Ann Arbor, MI 48109 USA

来源：

ELECTROPHORESIS | 2014年 / 35卷 / 15期

基金：

美国国家卫生研究院;

关键词：

Alpha-2-macroglobulin; Core-fucosylation; Label-free quantification; LC-MS/MS; Site-specific glycosylation; MASS-SPECTROMETRIC ANALYSIS; N-GLYCOSYLATION SITES; ENZYMATIC DEGLYCOSYLATION; GLYCOPEPTIDE ANALYSIS; SERUM; IDENTIFICATION; GLYCOPROTEINS; BIOMARKER; GLYCANS; CHROMATOGRAPHY;

D O I：

10.1002/elps.201300376

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a label-free relative quantification LC-MS/MSmethod for core-fucosylation in alpha-2-macroglobulin (A2MG) immunoprecipitated from human sera. The method utilizes endoglycosidase F partial deglycosylation to reduce glycosylation microheterogeneity, while retaining the innermost N-acetylglucosamine (GlcNAc) and core fucose. Precursor ion peak areas of partially deglycosylated peptides were obtained and site-specific core-fucosylation ratios based on the peak areas of core-fucosylated and nonfucosylated counterparts were calculated and evaluated for assay development. This assay was applied in a preliminary study of sera samples from normal controls and patients with pancreatic diseases, including pancreatic cancer and chronic pancreatitis. A2MG fucosylation levels at sites N396 and N1424 were found to decrease in both chronic pancreatitis and pancreatic cancer compared to normal controls. The two sites were identified by two peptides and their core-fucosylation ratios were found to be internally consistent. This method provides a platform to quantify fucosylation levels and can be used to study site-specific core-fucosylation aberrations in other glycoproteins for other diseases.

引用

页码：2108 / 2115

页数：8

共 26 条

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