蛋白质组学应用：压力循环技术催进蛋白酶解能力

蛋白质组学应用：压力循环技术催进蛋白酶解能力

Application of Pressurized Solvents for Ultrafast Trypsin Hydrolysis in Proteomics:

Proteomics on the Fly

摘要:
一种在高压下迅速水解蛋白质的新方法: 利用压力循环技术范围为5-35 kpsi;从而协助蛋白质组学分析。压力循环技术成功在60sec内把单一蛋白质及复杂的蛋白质水解.然后利用反相液相色谱分析。本文中, 利用压力循环技术新方法进行Shewanella oneidensis 多肽水解与传统的“过夜”样品制备方法作对比. 新方法-压力循环的优势:包括简化样品处理过程，易于实施，实验中样品没有交叉污染，和成本效率高。

Abstract:

A new method for rapid proteolytic digestion of proteins under high pressure that uses pressure cycling technology in the range of 5-35 kpsi was demonstrated for proteomic analysis. Successful in-solution digestions of single proteins and complex protein mixtures were achieved in 60 s and then analyzed by reversed phase liquid chromatography-electrospray ionization ion trap-mass spectrometry. Method performance in terms of the number of Shewanella oneidensis peptides and proteins identified in a shotgun approach was evaluated relative to a traditional “overnight” sample preparation method. Advantages of the new method include greatly simplified sample processing, easy implementation, no cross contamination among samples, and cost effectiveness.

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