Selected Reaction Monitoring - SRM in Proteomics

SRM in Proteomics

SRM can be used for targeted quantitative proteomics by mass spectrometry. Following ionization in an electrospray source, a peptide precursor is first isolated to obtain a substantial ion population of mostly the intended species. This population is then fragmented to yield product ions whose signal abundances are indicative of the abundance of the peptide in the sample. This experiment is primarily performed on triple quadrupole mass spectrometers, where mass-resolving Q₁ isolates the precursor, q₂ acts as a collision cell, and mass-resolving Q₃ is cycled through the product ions which are detected upon exiting the last quadrupole by an electron multiplier. A precursor/product pair is often referred to as a transition. Much work goes into ensuring that transitions are selected that have maximum specificity. By spiking in heavy-labeled (e.g., D, 13C, or 15N) peptides to a complex matrix as concentration standards, SRM can be used to construct a calibration curve that can provide the absolute quantification (i.e., copy number per cell) of the native, light peptide, and by extension, its parent protein.