Figure 1. Protein identification by mass spectrometry.
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Figure 2. Structure of the 4‐plex iTRAQ reagent. The reagent consists of an amine‐reactive group, a balance group, and a reporter group. The isobaric tags have a mass of 145 Da, which results in a mass difference of 144.1 Da for all differently labeled peptides. Upon fragmentation, the reporter group and a neutral fragment (balance group) are released. Due to the isotope composition of the reporter and the balance group, the generated reporter ions show different masses for the different iTRAQ reagents.
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Figure 3. MS‐based quantification of spliceosomal B and C complexes. (a) MS spectrum of peptides derived from metabolically (SILAC) labeled B and C complexes. C complexes were purified from a light nuclear extract, and B complexes from a heavy nuclear extract. The signal intensities of the differently labeled peptides are used for quantification; (b) MS/MS spectrum of chemically (iTRAQ) labeled peptides derived from B and C complexes. The signal intensities of the iTRAQ reporter ions are used for quantification. Peptides derived from B complexes were labeled with iTRAQ‐115, and peptides derived from C complexes with iTRAQ‐116. The internal standard (containing a mixture of B and C complex peptides) was labeled with iTRAQ‐114.
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