Evaluation of a High-Intensity Focused Ultrasound-Immobilized Trypsin Digestion and 18O-Labeling Method for Quantitative Proteomics

被引：30

作者：

Lopez-Ferrer, Daniel ^{[1
]}

Hixson, Kim K. ^{[2
]}

Smallwood, Heather ^{[1
]}

Squier, Thomas C. ^{[1
]}

Petritis, Konstantinos ^{[1
]}

Smith, Richard D. ^{[1
]}

机构：

[1] Pacific NW Natl Lab, Div Biol Sci, Richland, WA 99352 USA

[2] Pacific NW Natl Lab, Environm Mol Sci Lab, Richland, WA 99352 USA

来源：

ANALYTICAL CHEMISTRY | 2009年 / 81卷 / 15期

关键词：

MASS-SPECTROMETRY; LIQUID-CHROMATOGRAPHY; PROTEINS; IDENTIFICATION; ENHANCEMENT; EXCHANGE; MODEL;

D O I：

10.1021/ac802540s

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

A new method that uses immobilized trypsin concomitant with ultrasonic irradiation results in ultrarapid digestion and more thorough O-18 labeling for quantitative protein comparisons. The method was reproducible and provided effective digestions within < 1 min with lower amounts of enzyme, compared to traditional methods. This method was demonstrated for digestion of both simple and complex protein mixtures, including bovine serum albumin, a global proteome extract from the bacteria Shewanella oneidensis, and mouse plasma, as well as O-18 labeling of complex protein mixtures, validating this method for differential proteomic measurements. This approach is simple, reproducible, cost-effective, rapid, and well suited for automation.

引用

页码：6272 / 6277

页数：6

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