Accurate Mass Multiplexed Tandem Mass Spectrometry for High-Throughput Polypeptide Identification from Mixtures
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- 作者：Christophe Masselon ; Gordon A. Anderson ; Richard Harkewicz ; James E. Bruce ; Ljiljana Pasa-Tolic ; and Richard D. Smith
- 刊名：Analytical Chemistry
- 出版年：2000
- 出版时间：April 15, 2000
- 年：2000
- 卷：72
- 期：8
- 页码：1918 - 1924
- 全文大小：130K
- 年卷期：v.72,no.8(April 15, 2000)
- ISSN：1520-6882
文摘
We report a new tandem mass spectrometric approachfor the improved identification of polypeptides frommixtures (e.g., using genomic databases). The approachinvolves the dissociation of several species simultaneouslyin a single experiment and provides both increased speedand sensitivity. The data analysis makes use of the knownfragmentation pathways for polypeptides and highly accurate mass measurements for both the set of parentpolypeptides and their fragments. The accurate massinformation makes it possible to attribute most fragmentsto a specific parent species. We provide an initial demonstration of this multiplexed tandem MS approach usingan FTICR mass spectrometer with a mixture of sevenpolypeptides dissociated using infrared irradiation froma CO2 laser. The peptides were added to, and thensuccessfully identified from, the largest genomic databaseyet available (C. elegans), which is equivalent in complexity to that for a specific differentiated mammalian celltype. Additionally, since only a few enzymatic fragmentsare necessary to unambiguously identify a protein froman appropriate database, it is anticipated that the multiplexed MS/MS method will allow the more rapid identification of complex protein mixtures with on-line separation of their enzymatically produced polypeptides.