Quantitative analysis of acrylamide labeled serum proteins by LC-MS/MS

被引：92

作者：

Faca, Vitor ^{[1
]}

Coram, Marc ^{[1
]}

Phanstiel, Doug ^{[1
]}

Glukhova, Veronika ^{[1
]}

Zhang, Qing ^{[1
]}

Fitzgibbon, Matthew ^{[1
]}

McIntosh, Martin ^{[1
]}

Hanash, Samir ^{[1
]}

机构：

[1] Fred Hutchinson Canc Res Ctr, Seattle, WA 98109 USA

来源：

JOURNAL OF PROTEOME RESEARCH | 2006年 / 5卷 / 08期

关键词：

acrylamide isotope labeling; LTQ-FTICR; human serum;

D O I：

10.1021/pr060102+

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

Isotopic labeling of cysteine residues with acrylamide was previously utilized for relative quantitation of proteins by MALDI-TOF. Here, we explored and compared the application of deuterated and C-13 isotopes of acrylamide for quantitative proteomic analysis using LC-MS/MS and high-resolution FTICR mass spectrometry. The method was applied to human serum samples that were immunodepleted of abundant proteins. Our results show reliable quantitation of proteins across an abundance range that spans 5 orders of magnitude based on ion intensities and known protein concentration in plasma. The use of C-13 isotope of acrylamide had a slightly greater advantage relative to deuterated acrylamide, because of shifts in elution of deuterated acrylamide relative to its corresponding nondeuterated compound by reversed-phase chromatography. Overall, the use of acrylamide for differentially labeling intact proteins in complex mixtures, in combination with LC-MS/MS provides a robust method for quantitative analysis of complex proteomes.

引用

页码：2009 / 2018

页数：10

共 19 条

[1] Selective detection of membrane proteins without antibodies - A mass spectrometric version of the Western blot [J].