蛋白質(zhì)組學(xué)應(yīng)用：壓力循環(huán)技術(shù)催進(jìn)蛋白酶解能力

Application of Pressurized Solvents for Ultrafast Trypsin Hydrolysis in Proteomics:

Proteomics on the Fly
 

摘要:
    一種在高壓下迅速水解蛋白質(zhì)的新方法: 利用壓力循環(huán)技術(shù)范圍為5-35 kpsi;從而協(xié)助蛋白質(zhì)組學(xué)分析。壓力循環(huán)技術(shù)成功在60sec內(nèi)把單一蛋白質(zhì)及復(fù)雜的蛋白質(zhì)水解.然后利用反相液相色譜分析。本文中, 利用壓力循環(huán)技術(shù)新方法進(jìn)行Shewanella oneidensis 多肽水解與傳統(tǒng)的“過(guò)夜”樣品制備方法作對(duì)比.  新方法-壓力循環(huán)的優(yōu)勢(shì):包括簡(jiǎn)化樣品處理過(guò)程，易于實(shí)施，實(shí)驗(yàn)中樣品沒有交叉污染，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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