Mass and Composition Matrix-Assisted Laser Desorption Ionization Time of Flight Analysis Enabling Inference of the Sequence of Most Peptides Where the Protein of Origin Is Known

Mass spectrometers combining matrix-assisted laser-desorption ionization (MALDI) and time-of-flight analysis are among the most widely used in peptide analysis. They excel at accurate mass determinations on complex samples but, compared with tandem instruments, have very limited capacity to determine amino acid sequence through daughter ion analysis. Here we have investigated the sequence information that can be inferred from the masses of peptides in the special circumstance in which the peptides are known to be sub-sequences of known parent sequences. We show how sequence can be inferred from the measured m/z of a peptide (mass analysis) and examine the parameters that influence the level of confidence that can be placed in "inferred sequences". We further describe how specific amino acid modifications can be used with MALDI-TOF analysis to obtain partial composition information and demonstrate that combined mass and composition (MAC) analysis enables the sequences of most peptide ions to be inferred with very high confidence.