MS/MS analysis of a cross-linked product derived from a GluC-digest of PKG, which was cross-linked with W45-[Phe(Bz)-6]. A, the [M+5H]5+ precursor ion at m/z 590–593 that was selected by the quadrupole using a broad selection window (∼5 m/z units). The characteristic doublet is indicative for the presence of the isotopically labeled leucine from the photoaffinity labeled peptide. B, MS/MS spectrum of the [M+5H]5+ at m/z 590–593. C, primary sequence of the cross-linked product and identified fragment ions. The b and y″ ions corresponding the primary sequence of PKG, without covalently attached inhibitor are annotated by bn or yn, whereas b or y″ ions of the primary sequence of PKG that carries the covalent attached photolabel are labeled *bn or *yn.