Hixson Kim K
Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA, USA.
Methods Mol Biol. 2009;492:39-63. doi: 10.1007/978-1-59745-493-3_3.
Prokaryotic protein expression changes in detectable amounts due to the environmental stimuli encountered by the organism. To understand the underlying mechanisms involved it is necessary to comprehensively detect both the proteins present and their relative abundance under the growth conditions of interest. LC-MS based accurate mass and time (AMT) tag method along with the use of clustering software can provide a visual and more comprehensive understanding of significant protein abundance increases and decreases. These data then can be effectively used to pin-point proteins of interest for further genetic and physiological studies. This method allows for the identification and quantitation of thousands of proteins in a single mass spectrometric analysis and is more comprehensive than two dimensional electrophoresis and shotgun approaches.
由于生物体遇到的环境刺激,原核生物的蛋白质表达会发生可检测量的变化。为了理解其中的潜在机制,有必要在感兴趣的生长条件下全面检测存在的蛋白质及其相对丰度。基于液相色谱-质谱联用的精确质量和时间(AMT)标签法以及聚类软件的使用,可以对蛋白质丰度的显著增加和减少提供直观且更全面的理解。然后,这些数据可以有效地用于确定感兴趣的蛋白质,以便进行进一步的遗传和生理学研究。这种方法允许在单次质谱分析中鉴定和定量数千种蛋白质,并且比二维电泳和鸟枪法更全面。