Solid Phase Isobaric Mass Tag Reagent for Simultaneous Protein Identification and Assay

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• 作者：Anna Napoli ; Constantinos M. Athanassopoulos ; Petros Moschidis ; Donatella Aiello ; Leonardo Di Donna ; Fabio Mazzotti ; Giovanni Sindona
• 刊名：Analytical Chemistry
• 出版年：2010
• 出版时间：July 1, 2010
• 年：2010
• 卷：82
• 期：13
• 页码：5552-5560
• 全文大小：332K
• 年卷期：v.82,no.13(July 1, 2010)
• ISSN：1520-6882

文摘

The solid phase isobaric mass tagging (SPIMT) approach is presented for simultaneous protein quantitation and identification. The novelty of the SPIMT strategy relies on a CID-based differentiation of regioisomeric species for quantitation of tagged proteolytic peptides. SPIMTs are unlabeled mass-tagging reagents, which consist of a reporter group, a mass balance group, and a spacer with a amine-specific reactive group, able to be linked to any N-terminal peptide. Therefore SPIMT-linked peptides from a two-plex set appear as a single unresolved precursor ion in MS, whereas the reporter groups lead to quantitation signals of m/z 168.2 and 182.2 Da upon tandem mass spectrometry (MS/MS) analysis with matrix-assisted laser desorption time-of-flight/time-of-flight (MALDI TOF/TOF). This strategy allows ease protein identification by direct submission of MS and MS/MS data to the MASCOT database. SPIMT approach showed an excellent quantitation linearity, detecting any relative concentration differences of peptides in two solutions over a 5-fold concentration range without losing sequencing information. Therefore, SPIMTs are an attractive, simple, and low cost alternative for two-plex quantitation of proteins and offer possibilities of tuning the two-plex signal mass window by replacing the spacer.