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Tandem MS (MS/MS) on the Q-ToF2 MS2MS1. On-line LC-MSMS on the Q-ToF2 : peptides from a single protein Chromatogram Fragment Mass spectrum (MSMS): fragments.

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Presentation on theme: "Tandem MS (MS/MS) on the Q-ToF2 MS2MS1. On-line LC-MSMS on the Q-ToF2 : peptides from a single protein Chromatogram Fragment Mass spectrum (MSMS): fragments."— Presentation transcript:

1 Tandem MS (MS/MS) on the Q-ToF2 MS2MS1

2 On-line LC-MSMS on the Q-ToF2 : peptides from a single protein Chromatogram Fragment Mass spectrum (MSMS): fragments from one peptide time Separate peptides over a 60 minute LC run, perform tandem MS on each peptide Survey Mass Spectrum - intact peptides mass/charge

3 Tandem MS - peptide de novo sequencing series of peptide fragments each fragment is one amino acid longer than the next the series of fragments corresponds to the sequence of the peptide low energy collision fragments the peptide cleavage usually occurs at the amide bond i.e. between residues

4 peptide de novo sequencing the series of fragments corresponds to the sequence of the peptide

5 De novo sequencing Sequence reads in N to C direction - PSGASTGVHEAMR

6 tandem MS / de novo sequences used for database searches for protein identification eg BLAST confirm sequence of engineered mutations, alleles, etc examine modifications – biotinylation, acetylation, oxidation, phosphorylation, glycosylation peptide de novo sequencing

7 Methionine removed Serine acetylated Modifications e.g. N-terminus S-acetyl


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