Aggarwal Kunal, Choe Leila H, Lee Kelvin H
School of Chemical and Biomolecular Engineering, Cornell University, 120 Olin Hall, Ithaca, NY 14853-5201, USA.
Brief Funct Genomic Proteomic. 2006 Jun;5(2):112-20. doi: 10.1093/bfgp/ell018. Epub 2006 May 10.
Shotgun proteomic methods involving isobaric tagging of peptides enable high-throughput proteomic analysis. iTRAQ reagents allow simultaneous identification and quantitation of proteins in four different samples using tandem mass spectrometry (MS). In this article, we provide a brief description of proteome analysis using iTRAQ reagents and review the current applications of these reagents in proteomic studies. We also compare different aspects of protein identification including protein sequence coverage and proteome coverage obtained using iTRAQ reagents with those using other shotgun proteomic techniques. We briefly discuss the issue of isotope purity correction in measured peak areas during protein quantitation using iTRAQ reagents. Finally, we conclude with some of the current challenges in MS-based proteomic analysis that are limiting protein identifications obtained by different shotgun proteomic methods.
涉及肽段等压标记的鸟枪法蛋白质组学方法能够实现高通量蛋白质组分析。iTRAQ试剂可利用串联质谱（MS）同时鉴定和定量四个不同样品中的蛋白质。在本文中，我们简要描述了使用iTRAQ试剂进行蛋白质组分析的过程，并综述了这些试剂在蛋白质组学研究中的当前应用。我们还比较了蛋白质鉴定的不同方面，包括使用iTRAQ试剂获得的蛋白质序列覆盖率和蛋白质组覆盖率与使用其他鸟枪法蛋白质组学技术获得的情况。我们简要讨论了在使用iTRAQ试剂进行蛋白质定量时测量峰面积中的同位素纯度校正问题。最后，我们总结了基于质谱的蛋白质组分析中目前存在的一些挑战，这些挑战限制了通过不同鸟枪法蛋白质组学方法获得的蛋白质鉴定。
