Practice question
Question
A shift of 98 Da less than expected in MS/MS means:
Explanation
Phosphorylation adds 80 Da, but under collision-induced dissociation phosphorylated serine and threonine frequently undergo beta-elimination of phosphoric acid H3PO4 with mass 97.9769 Da, rounded to 98 Da loss from precursor or product ions. Observation of peak 98 Da lower than predicted molecular weight signals neutral loss of phosphate, indicating presence of phosphopeptide that lost H3PO4 during fragmentation. This diagnostic loss distinguishes phosphorylated peptides from unmodified counterparts and contrasts with oxidation or deamination shifts. Recognition of -98 Da loss supports phosphosite localization in signal transduction studies and proteomics research widely used.