Peptide photodissociation with 157 nm light in a commercial tandem time-of-flight mass spectrometer.

Photodissociation with 157 nm light was implemented in an ABI model 4700 matrix-assisted laser desorption ionization (MALDI) tandem time-of-flight (TOF) mass spectrometer for peptide analysis. With a homemade computer program to control the light timing based on the m/z of each precursor ion, the photodissociation setup was seamlessly automated with the mass spectrometer. Peptide photodissociation in this apparatus yielded fragments similar to those observed in previous experiments with a home-built tandem-TOF mass spectrometer. Peptides having arginine at their C-termini yielded high-energy x-, v-, and w- type fragments, while peptides with N-terminal arginine produced many a- and d- type ions. Abundant immonium ions were also generated. High-quality photodissociation spectra were obtained with as little as 5 fmol of peptides. In the analysis of various tryptic peptides, photodissociation provided much more sequence information than the conventional TOF-TOF collision induced dissociation (CID). Because of the high fragmentation efficiency, sensitivity was not sacrificed to achieve this.