Relative stability of peptide sequence ions generated by tandem mass spectrometry.

We report the use of unimolecular dissociation by infrared radiation for gaseous multiphoton energy transfer to determine relative activation energy (E(a,laser)) for dissociation of peptide sequence ions. The sequence ions of interest are mass-isolated; the entire ion cloud is then irradiated with a continuous wave CO(2) laser, and the first order rate constant, k(d), is determined for each of a series of laser powers. Provided these conditions are met, a plot of the natural logarithm of k(d) versus the natural logarithm of laser power yields a straight line, whose slope provides a measure of E(a,laser). This method reproduces the E(a) values from blackbody radiative dissociation (BIRD) for the comparatively large, singly and doubly protonated bradykinin ions (nominally y ( 9 ) and y ( 9 ) ( 2+ )). The comparatively small sequence ion systems produce E(a,laser) values that are systematic underestimates of theoretical barriers calculated with density functional theory (DFT). However, the relative E(a,laser) values are in qualitative agreement with the mobile proton model and available theory. Additionally, novel protonated cyclic-dipeptide (diketopiperazine) fragmentation reactions are analyzed with DFT. FT-ICR MS provides access to sequence ions generated by electron capture dissociation, infrared multiphoton dissociation, and collisional activation methods (i.e., b ( n ), y ( m ) , c ( n ), z ( m ) ( • ) ions).